Nguyễn tài sức triển khai mô hình thực nghiệm gây ung thư não trên chuột suy giảm miễn dịch khóa luận tốt nghiệp dược sĩ

BỘ Y TẾ TRƯỜNG ĐẠI HỌC DƯỢC HÀ NỘI - - NGUYỄN TÀI SỨC TRIỂN KHAI MÔ HÌNH THỰC NGHIỆM GÂY UNG THƯ NÃO TRÊN CHUỘT SUY GIẢM MIỄN DỊCH KHÓA LUẬN TỐT NGHIỆP DƯỢC SĨ HÀ NỘI – 2023 BỘ Y TẾ TRƯỜNG ĐẠI HỌC DƯỢC HÀ NỘI - - NGUYỄN TÀI SỨC MÃ SINH VIÊN: 1801608 TRIỂN KHAI MƠ HÌNH THỰC NGHIỆM GÂY UNG THƯ NÃO TRÊN CHUỘT SUY GIẢM MIỄN DỊCH KHÓA LUẬN TỐT NGHIỆP DƯỢC SĨ Người hướng dẫn: PGS.TS Nguyễn Thị Lập TS BS Bùi Khắc Cường Nơi thực hiện: Trung tâm Nghiên cứu động vật thực nghiệm Học viện Quân y Trường Đại học Dược Hà Nội HÀ NỘI - 2023 LỜI CẢM ƠN Đầu tiên, xin gửi lời cảm ơn chân thành sâu sắc đến PGS.TS Nguyễn Thị Lập, Bộ mơn Hóa sinh, Trường Đại học Dược Hà Nội, nhận hướng dẫn tạo điều kiện tốt cho tơi hồn thành khóa luận Đặc biệt, xin gửi lời cảm ơn chân thành sâu sắc đến TS.BS Bùi Khắc Cường, Phó giám đốc Trung tâm nghiên cứu động vật thực nghiệm, Học viện Qn y, nhận hướng dẫn tơi tạo điều kiện tốt cho tơi hồn thành khóa luận Kế đến, xin gửi lời cảm ơn chân thành sâu sắc đến anh chị làm việc Trung tâm Nghiên cứu động vật thực nghiệm, Học viện Quân y Khoa Giải phẫu bệnh-Pháp y, Bệnh viện Quân y 103 hướng dẫn tận tình tạo điều kiện tốt cho khoảng thời gian làm thực nghiệm Nhân đây, xin gửi lời cảm ơn đến Ban Giám hiệu tồn thể thầy Trường Đại học Dược Hà Nội giảng dạy tận tình tạo điều kiện khoảng thời gian tơi học tập trường để tơi có tảng kiến thức để hồn thành khóa luận hành nghề sau Cuối cùng, xin gửi lời cảm ơn đến gia đình bạn bè tơi động viên, chia sẻ, ủng hộ chỗ dựa tinh thần vững đường nghiên cứu khoa học Do thời gian làm thực nghiệm có giới hạn kiến thức thân cịn hạn chế, nên khóa luận khơng tránh khỏi nhiều thiếu sót Tơi mong nhận góp ý thầy cơ, bạn bè để khóa luận hồn thiện Tơi xin chân thành cảm ơn! Hà Nội, ngày 05 tháng 06 năm 2023 Sinh viên Nguyễn Tài Sức MỤC LỤC DANH MỤC CÁC CHỮ VIẾT TẮT, KÝ HIỆU DANH MỤC CÁC BẢNG DANH MỤC CÁC HÌNH VẼ ĐẶT VẤN ĐỀ CHƯƠNG TỔNG QUAN 1.1 Tổng quan u nguyên bào thần kinh đệm ác tính, ung thư đại trực tràng ung thư đại trực tràng di não 1.1.1 Đặc điểm dịch tễ 1.1.2 Các yếu tố nguy 1.1.3 Cơ chế bệnh sinh ung thư 1.2 Tổng quan mơ hình u não chuột 10 1.2.1 Mô hình đồng ghép 11 1.2.2 Mơ hình động vật biến đổi gen 12 1.2.3 Mơ hình dựa dòng tế bào ung thư người 12 1.2.4 Mơ hình dựa mẫu bệnh phẩm bệnh nhân 13 CHƯƠNG ĐỐI TƯỢNG VÀ PHƯƠNG PHÁP NGHIÊN CỨU 14 2.1 Đối tượng nghiên cứu 14 2.1.1 Động vật dùng nghiên cứu 14 2.1.2 Dòng tế bào dùng nghiên cứu 14 2.2 Nguyên liệu trang thiết bị dùng nghiên cứu 14 2.2.1 Trang thiết bị, dụng cụ dùng nghiên cứu 14 2.2.2 Hoá chất dùng nghiên cứu 15 2.3 Phương pháp nghiên cứu 16 2.3.1 Ni cấy tăng sinh dịng tế bào ung thư HCT116 U87MG 16 2.3.2 Tạo mơ hình u não chuột 17 2.3.3 Xử lý số liệu 20 2.3.4 Địa điểm nghiên cứu 20 2.3.5 Sơ đồ nghiên cứu 21 CHƯƠNG KẾT QUẢ NGHIÊN CỨU 22 3.1 Kết ni cấy tăng sinh dịng tế bào HCT116 U87MG 22 3.2 Kết sau ghép dòng tế bào HCT116 U87MG vào não chuột C57BL/6 BALB/c gây suy giảm miễn dịch chiếu xạ 22 3.2.1 Kết mơ hình ghép dịng tế bào ung thư đại trực tràng người HCT116 vào não chuột nhắt chủng C57BL/6 22 3.2.2 Kết mơ hình ghép dòng tế bào ung thư đại trực tràng người HCT116 vào não chuột nhắt chủng BALB/c 26 3.2.3 Kết mơ hình ghép dịng tế bào u ngun bào thần kinh đệm U87MG vào não chuột nhắt chủng BALB/c 29 CHƯƠNG BÀN LUẬN 34 4.1 Về lựa chọn mơ hình nghiên cứu 34 4.2 Về kết mơ hình nghiên cứu 36 4.2.1 Quá trình ni cấy dịng tế bào ung thư đại trực tràng HCT116 dòng tế bào u nguyên bào thần kinh đệm U87MG in vitro 36 4.2.2 Mơ hình ghép dịng tế bào ung thư HCT116 não chuột chủng C57BL/6 36 4.2.3 Mơ hình ghép dịng tế bào ung thư HCT116 não chuột chủng BALB/c 37 4.2.4 Mô hình ghép dịng tế bào ung thư U87MG não chuột chủng BALB/c 38 KẾT LUẬN 41 TÀI LIỆU THAM KHẢO PHỤ LỤC DANH MỤC CÁC CHỮ VIẾT TẮT, KÝ HIỆU Viết đầy đủ theo tiếng Việt Chữ viết tắt Viết đầy đủ theo tiếng Anh Protein liên kết yếu tố khởi Eukaryotic translation initiation động dịch mã 4E sinh vật 4EBP1 factor 4E-binding protein nhân chuẩn Enzym protein kinase B AKT Protein kinase B Protein ức chế đa polyp APC Adenomatous polyposis coli tuyến Bộ sưu tập chủng giống Hoa ATCC American Type Culture Collection Kỳ Protein liên quan đến tự thực Atg13 Autophagy Related 13 bào 13 Yếu tố tăng trưởng nguyên bFGF Basic fibroblast growth factor bào sợi The Central Brain Tumor Registry of Cơ quan đăng ký khối u não CBTRUS the United States trung ương Hoa Kỳ Kinase phụ thuộc cyclin CDK Cyclin-dependent kinase Ung thư đại trực tràng CRC Colorectal cancer Môi trường DMEM nuôi cấy DMEM Dulbecco's Modified Eagle Medium tế bào Acid deoxyribonucleic DNA Deoxyribonucleic acid Yếu tố tăng trưởng biểu bì EGF Epidermal growth factor Hội chứng đa polyp gia đình FAP Familial adenomatous polyposis Huyết bào thai bò FBS Fetal bovine serum Formalin fixing and paraffine Cố định formalin vùi FFPE embedding paraffin U nguyên bào thần kinh đệm GBM Glioblastoma multiforme ác tính Guanosine diphosphat GDP Guanosine diphosphate Growth factor receptor-bound Protein liên kết thụ thể yếu Grb2 protein tố tăng trưởng Guanosine triphosphat GTP Guanosine triphosphate Hematoxylin Eosin H&E Hematoxylin and Eosin Yếu tố cảm ứng tình trạng HIF1/2 Hypoxia-Inducible Factor-1/2 thiếu oxy-1/2 Enzym isocitrate IDH Isocitrate dehydrogenase dehydrogenase Phạm vi tứ phân vị IQR Interquartile range Protein thuộc họ RAS KRAS Kirsten rat sarcoma virus Protein kinase kích MAPK ERK Mitogen-activated protein kinase hoạt mitogen i MEK1/2 MLH1 MSH2 Mitogen-activated protein kinase kinase-1/2 MutL protein homolog MutS homolog mTOR Mammalian target of rapamycin mTORC1/2 Mammalian target of rapamycin complex-1/2 PBS phosphate buffered saline PDGF PIP3 PKC Platelet-derived growth factor Platelet-derived growth factor receptor Phosphoinositide-dependent protein kinase-1/2 Phosphatidylinositol-4,5bisphosphate 3-kinase Phosphatidylinositol 4,5bisphosphate Phosphatidylinositol-3, 4, 5triphosphate Protein kinase C PPARγ Peroxisome proliferator-activated receptor-gamma PTEN Phosphatase and tensin homolog RAF Rapidly accelerated fibrosarcoma RAS Rat sarcoma virus Rb Retinoblastoma protein Rheb RTK SD SH2 SH3 Ras homolog enriched in brain Receptor tyrosine kinase Standard deviation Src Homology Src Homology PDGFR PDK1/2 PI3K PIP2 Sos Protein kinase hoạt hóa MAPK-1/2 Protein tương đồng MutL Chất tương đồng MutS Đích rapamycin động vật có vú Phức hợp đích rapamycin động vật có vú1/2 Nước muối sinh lý đệm phosphat Yếu tố tăng trưởng có nguồn gốc từ tiểu cầu Thụ thể yếu tố tăng trưởng có nguồn gốc từ tiểu cầu Protein kinase phụ thuộc vào phosphoinositide – 1/2 Enzym phosphatidylinositol4,5-bisphosphat 3-kinase Phosphatidylinositol 4,5bisphosphat Phosphatidylinositol-3, 4, 5triphosphat Enzym protein kinase C Thụ thể kích hoạt chất tăng sinh peroxisomegamma Phosphatase chất tương đồng tensin Một loại protein kinase hoạt hóa MEK Một họ protein GTPase gây ung thư Protein u nguyên bào võng mạc Chất tương đồng Ras làm giàu não Thụ thể tyrosine kinase Độ lệch chuẩn Một yếu tố trao đổi guanin nucleotide điều hòa hoạt Son of Sevenless ii STAT3 Sterol regulatory element-binding transcription factor Signal transducer and activator of transcription TERT Telomerase reverse transcriptase TGF-α TSC1/2 uPAR Transforming growth factor alpha Tuberous sclerosis complex-1/2 Urokinase-type plasminogen activator receptor VEGF WHO Vascular endothelial growth factor World Health Organization SREBP1 iii động họ protein RAS Yếu tố phiên mã liên kết yếu tố điều hịa sterol Chất truyền tín hiệu kích hoạt phiên mã Enzyme phiên mã ngược telomerase Yếu tố tăng trưởng biến đổialpha Phức hợp xơ cứng củ-1/2 Thụ thể bề mặt hoạt hóa plasminogen urokinase Yếu tố tăng trưởng nội mô mạch máu Tổ chức Y tế Thế giới DANH MỤC CÁC BẢNG Bảng Kết phát triển khối u não nhóm chuột C57BL/6 đối chứng, ghép dòng tế bào HCT116 23 Bảng Kết phát triển khối u não nhóm chuột C57BL/6 chiếu xạ, ghép dịng tế bào HCT116 24 Bảng 3 Thể tích khối u nhóm chuột C57BL/6 đối chứng chiếu xạ, ghép dòng tế bào HCT116 25 Bảng Kết phát triển u não chuột BALB/c đối chứng, ghép dòng tế bào HCT116 26 Bảng Kết phát triển u não chuột BALB/c chiếu xạ, ghép dòng tế bào HCT116 27 Bảng Thể tích khối u nhóm chuột BALB/c đối chứng chiếu xạ, ghép dòng tế bào HCT116 28 Bảng Kết phát triển u não chuột BALB/c đối chứng, ghép dòng tế bào U87MG 30 Bảng Kết phát triển u não chuột BALB/c chiếu xạ, ghép dòng tế bào U87MG 31 Bảng Thể tích khối u nhóm chuột BALB/c đối chứng chiếu xạ, ghép dòng tế bào U87MG 32 iv DANH MỤC CÁC HÌNH VẼ Hình 1 Biểu đồ biểu diễn tỷ lệ mắc GBM (theo định nghĩa cũ WHO) (màu xanh) tỷ lệ mắc năm (màu cam) theo nhóm tuổi năm từ 2014-2018 Hình Con đường RAS/RAF/MAPK, đường PI3K/AKT/mTOR đường p16/Rb chế bệnh sinh GBM Hình Ví dụ minh họa chế bệnh sinh dẫn tới ung thư đại trực tràng người 10 Hình Chuột nhắt chủng C57BL/6 chuột nhắt trắng chủng BALB/c ni chăm sóc Trung tâm nghiên cứu động vật thí nghiệm, Học viện Quân y 14 Hình 2 Một số dụng cụ dùng trình ghép tế bào ung thư vào não chuột 19 Hình Hình ảnh tế bào HCT116 quan sát kính hiển vi quang học 22 Hình Hình ảnh tế bào U87MG quan sát kính hiển vi quang học 22 Hình 3 Biểu đồ so sánh kích thước khối u não chuột C57BL/6 ghép dịng tế bào HCT116 nhóm đối chứng nhóm chiếu xạ 25 Hình Biểu đồ so sánh kích thước khối u nhóm đối chứng nhóm chiếu xạ chuột BALB/c ghép dòng tế bào HCT116 29 Hình Biểu đồ thể thay đổi số cân nặng trung bình nhóm đối chứng nhóm chiếu xạ chuột BALB/c ghép dòng tế bào HCT116 thời gian theo dõi 29 Hình Biểu đồ so sánh kích thước khối u nhóm đối chứng nhóm chiếu xạ chuột BALB/c ghép dòng tế bào U87MG 33 Hình Biểu đồ thể thay đổi số cân nặng trung bình nhóm đối chứng nhóm chiếu xạ chuột BALB/c ghép dòng tế bào U87MG thời gian theo dõi 33 v TÀI LIỆU THAM KHẢO Tiếng Việt [1] Trần Minh Thông (2007), Đặc điểm giải phẫu bệnh 1187 ca u bào, Tạp chí Y học thành phố Hồ Chí Minh, Thành phố Hồ Chí Minh, tr.41 [2] Bùi Khắc Cường, Hồ Anh Sơn, Nguyễn Lĩnh Toàn (2012), Nghiên cứu tạo khối ung thư đại tràng người chuột thiếu hụt miễn dịch kỹ thuật ghép dị lồi, Tạp chí Y-Dược học Qn sự, Hà Nội Tiếng Anh [3] Q T Ostrom et al., “CBTRUS Statistical Report: Primary Brain and Central Nervous System Tumors Diagnosed in the United States in 2006-2010,” Neuro-Oncol., vol 15, no suppl 2, pp ii1–ii56, Nov 2013, doi: 10.1093/neuonc/not151 [4] Q T Ostrom et al., “CBTRUS Statistical Report: Primary brain and other central nervous system tumors diagnosed in the United States in 2010–2014,” Neuro-Oncol., vol 19, no suppl_5, pp v1–v88, Nov 2017, doi: 10.1093/neuonc/nox158 [5] Q T Ostrom, G Cioffi, K Waite, C Kruchko, and J S Barnholtz-Sloan, “CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2014–2018,” Neuro-Oncol., vol 23, no Supplement_3, pp iii1–iii105, Oct 2021, doi: 10.1093/neuonc/noab200 [6] S Müller et al., “Brain Metastases from Colorectal Cancer: A Systematic Review of the Literature and Meta-Analysis to Establish a Guideline for Daily Treatment,” Cancers, vol 13, no 4, p 900, Feb 2021, doi: 10.3390/cancers13040900 [7] N R C (US) C on I C Rodents, “Maintenance of Rodents Requiring Isolation,” in Immunodeficient Rodents: A Guide to Their Immunobiology, Husbandry, and Use, National Academies Press (US), 1989 Accessed: Jun 05, 2023 [Online] Available: https://www.ncbi.nlm.nih.gov/books/NBK218201/ [8] K Kita et al., “In vivo imaging xenograft models for the evaluation of anti-brain tumor efficacy of targeted drugs,” Cancer Med., vol 6, no 12, pp 2972–2983, Dec 2017, doi: 10.1002/cam4.1255 [9] D N Louis et al., “The 2021 WHO Classification of Tumors of the Central Nervous System: a summary,” Neuro-Oncol., vol 23, no 8, pp 1231–1251, Aug 2021, doi: 10.1093/neuonc/noab106 [10] H.-G Wirsching, E Galanis, and M Weller, “Glioblastoma,” in Handbook of Clinical Neurology, Elsevier, 2016, pp 381–397 doi: 10.1016/B978-0-12-8029978.00023-2 [11] P Kleihues and H Ohgaki, “Phenotype vs Genotype in the Evolution of Astrocytic Brain Tumors,” Toxicol Pathol., vol 28, no 1, pp 164–170, Jan 2000, doi: 10.1177/019262330002800121 [12] P Kleihues and H Ohgaki, “Primary and secondary glioblastomas: From concept to clinicaldiagnosis,” Neuro-Oncol., vol 1, no 1, pp 44–51, Jan 1999, doi: 10.1093/neuonc/1.1.44 [13] S Grochans et al., “Epidemiology of Glioblastoma Multiforme–Literature Review,” Cancers, vol 14, no 10, p 2412, May 2022, doi: 10.3390/cancers14102412 [14] J T Low et al., “Primary brain and other central nervous system tumors in the United States (2014-2018): A summary of the CBTRUS statistical report for clinicians,” Neuro-Oncol Pract., vol 9, no 3, pp 165–182, May 2022, doi: 10.1093/nop/npac015 [15] “What Is Colorectal Cancer? | CDC.” https://www.cdc.gov/cancer/colorectal/basic_info/what-is-colorectal-cancer.htm (accessed Apr 11, 2023) [16] H Sung et al., “Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries,” CA Cancer J Clin., vol 71, no 3, pp 209–249, May 2021, doi: 10.3322/caac.21660 [17] W Street, “Colorectal Cancer Facts & Figures 2020-2022” [18] A G Zauber et al., “Colonoscopic Polypectomy and Long-Term Prevention of Colorectal-Cancer Deaths,” N Engl J Med., vol 366, no 8, pp 687–696, Feb 2012, doi: 10.1056/NEJMoa1100370 [19] S Cotton, L Sharp, and J Little, “The Adenoma-Carcinoma Sequence and Prospects for the Prevention of Colorectal Neoplasia,” Crit Rev Oncog., vol 7, no 5–6, pp 293–342, 1996, doi: 10.1615/CritRevOncog.v7.i5-6.10 [20] “Colorectal Cancer — Cancer Stat Facts.” https://seer.cancer.gov/statfacts/html/colorect.html (accessed Apr 12, 2023) [21] L A Gloeckler Ries, M E Reichman, D R Lewis, B F Hankey, and B K Edwards, “Cancer Survival and Incidence from the Surveillance, Epidemiology, and End Results (SEER) Program,” The Oncologist, vol 8, no 6, pp 541–552, Dec 2003, doi: 10.1634/theoncologist.8-6-541 [22] L G M van der Geest, J Lam-Boer, M Koopman, C Verhoef, M A G Elferink, and J H W de Wilt, “Nationwide trends in incidence, treatment and survival of colorectal cancer patients with synchronous metastases,” Clin Exp Metastasis, vol 32, no 5, pp 457–465, Jun 2015, doi: 10.1007/s10585-015-9719-0 [23] L H Biller and D Schrag, “Diagnosis and Treatment of Metastatic Colorectal Cancer: A Review,” JAMA, vol 325, no 7, p 669, Feb 2021, doi: 10.1001/jama.2021.0106 [24] L Nayak, E Q Lee, and P Y Wen, “Epidemiology of Brain Metastases,” Curr Oncol Rep., vol 14, no 1, pp 48–54, Feb 2012, doi: 10.1007/s11912-011-0203-y [25] S Sadetzki et al., “Description of selected characteristics of familial glioma patients – Results from the Gliogene Consortium,” Eur J Cancer, vol 49, no 6, pp 1335–1345, Apr 2013, doi: 10.1016/j.ejca.2012.11.009 [26] J S Nelson, C M Burchfiel, D Fekedulegn, and M E Andrew, “Potential risk factors for incident glioblastoma multiforme: the Honolulu Heart Program and Honolulu-Asia Aging Study,” J Neurooncol., vol 109, no 2, pp 315–321, Sep 2012, doi: 10.1007/s11060-012-0895-3 [27] S Yonehara et al., “Clinical and epidemiologic characteristics of first primary tumors of the central nervous system and related organs among atomic bomb survivors in Hiroshima and Nagasaki, 1958-1995,” Cancer, vol 101, no 7, pp 1644– 1654, Oct 2004, doi: 10.1002/cncr.20543 [28] M Hauptmann et al., “Brain cancer after radiation exposure from CT examinations of children and young adults: results from the EPI-CT cohort study,” Lancet Oncol., vol 24, no 1, pp 45–53, Jan 2023, doi: 10.1016/S1470-2045(22)00655-6 [29] J T Lowery et al., “Understanding the contribution of family history to colorectal cancer risk and its clinical implications: A state-of-the-science review: Family History and Colorectal Cancer,” Cancer, vol 122, no 17, pp 2633–2645, Sep 2016, doi: 10.1002/cncr.30080 [30] D E O’Sullivan et al., “Risk Factors for Early-Onset Colorectal Cancer: A Systematic Review and Meta-analysis,” Clin Gastroenterol Hepatol., vol 20, no 6, pp 1229-1240.e5, Jun 2022, doi: 10.1016/j.cgh.2021.01.037 [31] B Levin et al., “Screening and Surveillance for the Early Detection of Colorectal Cancer and Adenomatous Polyps, 2008: A Joint Guideline from the American Cancer Society, the US Multi-Society Task Force on Colorectal Cancer, and the American College of Radiology,” CA Cancer J Clin., vol 58, no 3, pp 130–160, May 2008, doi: 10.3322/CA.2007.0018 [32] B M Ryan et al., “An analysis of genetic factors related to risk of inflammatory bowel disease and colon cancer,” Cancer Epidemiol., vol 38, no 5, pp 583–590, Oct 2014, doi: 10.1016/j.canep.2014.07.003 [33] M W M D Lutgens, M G H van Oijen, G J M G van der Heijden, F P Vleggaar, P D Siersema, and B Oldenburg, “Declining Risk of Colorectal Cancer in Inflammatory Bowel Disease: An Updated Meta-analysis of Population-based Cohort Studies,” Inflamm Bowel Dis., vol 19, no 4, pp 789–799, Mar 2013, doi: 10.1097/MIB.0b013e31828029c0 [34] M Kyrgiou et al., “Adiposity and cancer at major anatomical sites: umbrella review of the literature,” BMJ, p j477, Feb 2017, doi: 10.1136/bmj.j477 [35] P Rawla, T Sunkara, and A Barsouk, “Epidemiology of colorectal cancer: incidence, mortality, survival, and risk factors,” Gastroenterol Rev., vol 14, no 2, pp 89–103, 2019, doi: 10.5114/pg.2018.81072 [36] A Karahalios et al., “Change in weight and waist circumference and risk of colorectal cancer: results from the Melbourne Collaborative Cohort Study,” BMC Cancer, vol 16, no 1, p 157, Dec 2016, doi: 10.1186/s12885-016-2144-1 [37] E Botteri, S Iodice, V Bagnardi, S Raimondi, A B Lowenfels, and P Maisonneuve, “Smoking and Colorectal Cancer: A Meta-analysis,” JAMA, vol 300, no 23, p 2765, Dec 2008, doi: 10.1001/jama.2008.839 [38] S Cai, Y Li, Y Ding, K Chen, and M Jin, “Alcohol drinking and the risk of colorectal cancer death: a meta-analysis,” Eur J Cancer Prev., vol 23, no 6, pp 532–539, Nov 2014, doi: 10.1097/CEJ.0000000000000076 [39] D S M Chan et al., “Red and Processed Meat and Colorectal Cancer Incidence: Meta-Analysis of Prospective Studies,” PLoS ONE, vol 6, no 6, p e20456, Jun 2011, doi: 10.1371/journal.pone.0020456 [40] A Lewandowska, G Rudzki, T Lewandowski, A Stryjkowska-Góra, and S Rudzki, “Risk Factors for the Diagnosis of Colorectal Cancer,” Cancer Control, vol 29, p 107327482110566, Nov 2022, doi: 10.1177/10732748211056692 [41] Y Esemen et al., “Molecular Pathogenesis of Glioblastoma in Adults and Future Perspectives: A Systematic Review,” Int J Mol Sci., vol 23, no 5, p 2607, Feb 2022, doi: 10.3390/ijms23052607 [42] S R Hubbard and J H Till, “Protein Tyrosine Kinase Structure and Function,” Annu Rev Biochem., vol 69, no 1, pp 373–398, Jun 2000, doi: 10.1146/annurev.biochem.69.1.373 [43] R P Agashe, S M Lippman, and R Kurzrock, “JAK: Not Just Another Kinase,” Mol Cancer Ther., vol 21, no 12, pp 1757–1764, Dec 2022, doi: 10.1158/15357163.MCT-22-0323 [44] R Trenker and N Jura, “Receptor tyrosine kinase activation: From the ligand perspective,” Curr Opin Cell Biol., vol 63, pp 174–185, Apr 2020, doi: 10.1016/j.ceb.2020.01.016 [45] M A Lemmon and J Schlessinger, “Cell Signaling by Receptor Tyrosine Kinases,” Cell, vol 141, no 7, pp 1117–1134, Jun 2010, doi: 10.1016/j.cell.2010.06.011 [46] U Degirmenci, M Wang, and J Hu, “Targeting Aberrant RAS/RAF/MEK/ERK Signaling for Cancer Therapy,” Cells, vol 9, no 1, p 198, Jan 2020, doi: 10.3390/cells9010198 [47] E Y Skolnik et al., “The SH2/SH3 domain-containing protein GRB2 interacts with tyrosine-phosphorylated IRS1 and Shc: implications for insulin control of ras signalling.,” EMBO J., vol 12, no 5, pp 1929–1936, May 1993, doi: 10.1002/j.14602075.1993.tb05842.x [48] D K Morrison, “MAP Kinase Pathways,” Cold Spring Harb Perspect Biol., vol 4, no 11, pp a011254–a011254, Nov 2012, doi: 10.1101/cshperspect.a011254 [49] M Bergoug, M Doudeau, F Godin, C Mosrin, B Vallée, and H Bénédetti, “Neurofibromin Structure, Functions and Regulation,” Cells, vol 9, no 11, p 2365, Oct 2020, doi: 10.3390/cells9112365 [50] C B Knobbe, J Reifenberger, and G Reifenberger, “Mutation analysis of the Ras pathway genes NRAS, HRAS, KRAS and BRAF in glioblastomas,” Acta Neuropathol (Berl.), vol 108, no 6, pp 467–470, Dec 2004, doi: 10.1007/s00401004-0929-9 [51] The Cancer Genome Atlas Research Network, “Comprehensive genomic characterization defines human glioblastoma genes and core pathways,” Nature, vol 455, no 7216, pp 1061–1068, Oct 2008, doi: 10.1038/nature07385 [52] C Xian J., “EGF family of growth factors: essential roles and functional redundancy in the nerve system,” Front Biosci., vol 9, no 1–3, p 85, 2004, doi: 10.2741/1210 [53] R Romano and C Bucci, “Role of EGFR in the Nervous System,” Cells, vol 9, no 8, p 1887, Aug 2020, doi: 10.3390/cells9081887 [54] C W Brennan et al., “The Somatic Genomic Landscape of Glioblastoma,” Cell, vol 155, no 2, pp 462–477, Oct 2013, doi: 10.1016/j.cell.2013.09.034 [55] M Chaffanet et al., “EGF receptor amplification and expression in human brain tumours,” Eur J Cancer, vol 28, no 1, pp 11–17, Jan 1992, doi: 10.1016/09598049(92)90374-B [56] K Watanabe, O Tachibana, K Sato, Y Yonekawa, P Kleihues, and H Ohgaki, “Overexpression of the EGF Receptor and p53 Mutations are Mutually Exclusive in the Evolution of Primary and Secondary Glioblastomas,” Brain Pathol., vol 6, no 3, pp 217–223, Jul 1996, doi: 10.1111/j.1750-3639.1996.tb00848.x [57] H K Gan, A N Cvrljevic, and T G Johns, “The epidermal growth factor receptor variant III (EGFRvIII): where wild things are altered,” FEBS J., vol 280, no 21, pp 5350–5370, Nov 2013, doi: 10.1111/febs.12393 [58] R Nishikawa et al., “A mutant epidermal growth factor receptor common in human glioma confers enhanced tumorigenicity.,” Proc Natl Acad Sci., vol 91, no 16, pp 7727–7731, Aug 1994, doi: 10.1073/pnas.91.16.7727 [59] H.-J S Huang et al., “The Enhanced Tumorigenic Activity of a Mutant Epidermal Growth Factor Receptor Common in Human Cancers Is Mediated by Threshold Levels of Constitutive Tyrosine Phosphorylation and Unattenuated Signaling,” J Biol Chem., vol 272, no 5, pp 2927–2935, Jan 1997, doi: 10.1074/jbc.272.5.2927 [60] P S Mischel and T F Cloughesy, “Targeted Molecular Therapy of GBM,” Brain Pathol., vol 13, no 1, pp 52–61, Apr 2006, doi: 10.1111/j.17503639.2003.tb00006.x [61] “Increased HOXA5 expression provides a selective advantage for gain of whole chromosome in IDH wild-type glioblastoma PMC.” https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5959235/ (accessed May 16, 2023) [62] S Noorolyai, N Shajari, E Baghbani, S Sadreddini, and B Baradaran, “The relation between PI3K/AKT signalling pathway and cancer,” Gene, vol 698, pp 120– 128, May 2019, doi: 10.1016/j.gene.2019.02.076 [63] K A McDowell, G J Riggins, and G L Gallia, “Targeting the AKT Pathway in Glioblastoma,” Curr Pharm Des., vol 17, no 23, pp 2411–2420, Aug 2011, doi: 10.2174/138161211797249224 [64] D D Sarbassov, D A Guertin, S M Ali, and D M Sabatini, “Phosphorylation and Regulation of Akt/PKB by the Rictor-mTOR Complex,” Science, vol 307, no 5712, pp 1098–1101, Feb 2005, doi: 10.1126/science.1106148 [65] K Inoki, Y Li, T Xu, and K.-L Guan, “Rheb GTPase is a direct target of TSC2 GAP activity and regulates mTOR signaling,” Genes Dev., vol 17, no 15, pp 1829– 1834, Aug 2003, doi: 10.1101/gad.1110003 [66] Q.-W Fan and W A Weiss, “Inhibition of PI3K-Akt-mTOR Signaling in Glioblastoma by mTORC1/2 Inhibitors,” in mTOR, T Weichhart, Ed., in Methods in Molecular Biology, vol 821 Totowa, NJ: Humana Press, 2012, pp 349–359 doi: 10.1007/978-1-61779-430-8_22 [67] J E Kim and J Chen, “Regulation of Peroxisome Proliferator–Activated Receptor-γ Activity by Mammalian Target of Rapamycin and Amino Acids in Adipogenesis,” Diabetes, vol 53, no 11, pp 2748–2756, Nov 2004, doi: 10.2337/diabetes.53.11.2748 [68] I Bakan and M Laplante, “Connecting mTORC1 signaling to SREBP-1 activation,” Curr Opin Lipidol., vol 23, no 3, pp 226–234, Jun 2012, doi: 10.1097/MOL.0b013e328352dd03 [69] M Laplante and D M Sabatini, “mTOR Signaling in Growth Control and Disease,” Cell, vol 149, no 2, pp 274–293, Apr 2012, doi: 10.1016/j.cell.2012.03.017 [70] S H Advani, “Targeting mTOR pathway: A new concept in cancer therapy,” Indian J Med Paediatr Oncol., vol 31, no 04, pp 132–136, Oct 2010, doi: 10.4103/0971-5851.76197 [71] A Anandharaj, S Cinghu, and W.-Y Park, “Rapamycin-mediated mTOR inhibition attenuates survivin and sensitizes glioblastoma cells to radiation therapy,” Acta Biochim Biophys Sin., vol 43, no 4, pp 292–300, Apr 2011, doi: 10.1093/abbs/gmr012 [72] J Ma et al., “Mammalian target of rapamycin regulates murine and human cell differentiation through STAT3/p63/Jagged/Notch cascade,” J Clin Invest., vol 120, no 1, pp 103–114, Jan 2010, doi: 10.1172/JCI37964 [73] P Rajan, D M Panchision, L F Newell, and R D G McKay, “BMPs signal alternately through a SMAD or FRAP–STAT pathway to regulate fate choice in CNS stem cells,” J Cell Biol., vol 161, no 5, pp 911–921, Jun 2003, doi: 10.1083/jcb.200211021 [74] K Yokogami, S Wakisaka, J Avruch, and S A Reeves, “Serine phosphorylation and maximal activation of STAT3 during CNTF signaling is mediated by the rapamycin target mTOR,” Curr Biol., vol 10, no 1, pp 47–50, Jan 2000, doi: 10.1016/S0960-9822(99)00268-7 [75] W Fu, X Hou, L Dong, and W Hou, “Roles of STAT3 in the pathogenesis and treatment of glioblastoma,” Front Cell Dev Biol., vol 11, p 1098482, Feb 2023, doi: 10.3389/fcell.2023.1098482 [76] S O Rahaman, P C Harbor, O Chernova, G H Barnett, M A Vogelbaum, and S J Haque, “Inhibition of constitutively active Stat3 suppresses proliferation and induces apoptosis in glioblastoma multiforme cells,” Oncogene, vol 21, no 55, pp 8404–8413, Dec 2002, doi: 10.1038/sj.onc.1206047 [77] S Zou, Q Tong, B Liu, W Huang, Y Tian, and X Fu, “Targeting STAT3 in Cancer Immunotherapy,” Mol Cancer, vol 19, no 1, p 145, Dec 2020, doi: 10.1186/s12943-020-01258-7 [78] M Abou-Ghazal et al., “The Incidence, Correlation with Tumor-Infiltrating Inflammation, and Prognosis of Phosphorylated STAT3 Expression in Human Gliomas,” Clin Cancer Res., vol 14, no 24, pp 8228–8235, Dec 2008, doi: 10.1158/1078-0432.CCR-08-1329 [79] K Masui, W K Cavenee, and P S Mischel, “mTORC2 in the center of cancer metabolic reprogramming,” Trends Endocrinol Metab., vol 25, no 7, pp 364–373, Jul 2014, doi: 10.1016/j.tem.2014.04.002 [80] W Fu and M N Hall, “Regulation of mTORC2 Signaling,” Genes, vol 11, no 9, p 1045, Sep 2020, doi: 10.3390/genes11091045 [81] C A Worby and J E Dixon, “PTEN,” Annu Rev Biochem., vol 83, no 1, pp 641–669, Jun 2014, doi: 10.1146/annurev-biochem-082411-113907 [82] C.-Y Chen, J Chen, L He, and B L Stiles, “PTEN: Tumor Suppressor and Metabolic Regulator,” Front Endocrinol., vol 9, p 338, Jul 2018, doi: 10.3389/fendo.2018.00338 [83] C Bassi et al., “Nuclear PTEN controls DNA repair and sensitivity to genotoxic stress,” Science, vol 341, no 6144, p 395, Jul 2013, doi: 10.1126/science.1236188 [84] P L Dahia, “PTEN, a unique tumor suppressor gene.,” Endocr Relat Cancer, pp 115–129, Jun 2000, doi: 10.1677/erc.0.0070115 [85] J M Trimarchi and J A Lees, “Sibling rivalry in the E2F family,” Nat Rev Mol Cell Biol., vol 3, no 1, pp 11–20, Jan 2002, doi: 10.1038/nrm714 [86] A Haque, N L Banik, and S K Ray, “Molecular Alterations in Glioblastoma,” in Progress in Molecular Biology and Translational Science, Elsevier, 2011, pp 187– 234 doi: 10.1016/B978-0-12-385506-0.00005-3 [87] K Reinhart et al., “Prevalence of flat lesions in a large screening population and their role in colonoscopy quality improvement,” Endoscopy, vol 45, no 05, pp 350– 356, Apr 2013, doi: 10.1055/s-0032-1326348 [88] D A Corley et al., “Variation of Adenoma Prevalence by Age, Sex, Race, and Colon Location in a Large Population: Implications for Screening and Quality Programs,” Clin Gastroenterol Hepatol., vol 11, no 2, pp 172–180, Feb 2013, doi: 10.1016/j.cgh.2012.09.010 [89] J S Levine and D J Ahnen, “Adenomatous Polyps of the Colon,” N Engl J Med., vol 355, no 24, pp 2551–2557, Dec 2006, doi: 10.1056/NEJMcp063038 [90] M Risio, “The natural history of adenomas,” Best Pract Res Clin Gastroenterol., vol 24, no 3, pp 271–280, Jun 2010, doi: 10.1016/j.bpg.2010.04.005 [91] E Dekker, P J Tanis, J L A Vleugels, P M Kasi, and M B Wallace, “Colorectal cancer,” The Lancet, vol 394, no 10207, pp 1467–1480, Oct 2019, doi: 10.1016/S0140-6736(19)32319-0 [92] “Gene expression analysis of colon high-grade dysplasia revealed new molecular mechanism of disease PMC.” https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6347998/ (accessed Apr 19, 2023) [93] S A Bien et al., “Genetic variant predictors of gene expression provide new insight into risk of colorectal cancer,” Hum Genet., vol 138, no 4, pp 307–326, Apr 2019, doi: 10.1007/s00439-019-01989-8 [94] E R Fearon and B Vogelstein, “A genetic model for colorectal tumorigenesis,” Cell, vol 61, no 5, pp 759–767, Jun 1990, doi: 10.1016/0092-8674(90)90186-i [95] K Motaparthi, S R Lauer, R M Patel, C I Vidal, and K Linos, “MYC gene amplification by fluorescence in situ hybridization and MYC protein expression by immunohistochemistry in the diagnosis of cutaneous angiosarcoma: Systematic review and appropriate use criteria,” J Cutan Pathol., vol 48, no 4, pp 578–586, Apr 2021, doi: 10.1111/cup.13912 [96] T A Baudino et al., “c-Myc is essential for vasculogenesis and angiogenesis during development and tumor progression,” Genes Dev., vol 16, no 19, pp 2530– 2543, Oct 2002, doi: 10.1101/gad.1024602 [97] M S Schrock, B R Stromberg, L Scarberry, and M K Summers, “APC/C ubiquitin ligase: Functions and mechanisms in tumorigenesis,” Semin Cancer Biol., vol 67, pp 80–91, Dec 2020, doi: 10.1016/j.semcancer.2020.03.001 [98] M Katoh, “Multi‑ layered prevention and treatment of chronic inflammation, organ fibrosis and cancer associated with canonical WNT/β‑ catenin signaling activation (Review),” Int J Mol Med., May 2018, doi: 10.3892/ijmm.2018.3689 [99] B Mann et al., “Target genes of β-catenin–T cell-factor/lymphoid-enhancer-factor signaling in human colorectal carcinomas,” Proc Natl Acad Sci., vol 96, no 4, pp 1603–1608, Feb 1999, doi: 10.1073/pnas.96.4.1603 [100] L Zhang and J W Shay, “Multiple Roles of APC and its Therapeutic Implications in Colorectal Cancer,” JNCI J Natl Cancer Inst., vol 109, no 8, Aug 2017, doi: 10.1093/jnci/djw332 [101] S M Powell et al., “APC mutations occur early during colorectal tumorigenesis,” Nature, vol 359, no 6392, pp 235–237, Sep 1992, doi: 10.1038/359235a0 [102] Y Rui, C Wang, Z Zhou, X Zhong, and Y Yu, “K-Ras mutation and prognosis of colorectal cancer: a meta-analysis,” Hepatogastroenterology., vol 62, no 137, pp 19– 24, 2015 [103] D Santini et al., “High Concordance of KRAS Status Between Primary Colorectal Tumors and Related Metastatic Sites: Implications for Clinical Practice,” The Oncologist, vol 13, no 12, pp 1270–1275, Dec 2008, doi: 10.1634/theoncologist.2008-0181 [104] “Targeting mutant KRAS - PubMed.” https://pubmed.ncbi.nlm.nih.gov/33838397/ (accessed Apr 22, 2023) [105] R Ullah, Q Yin, A H Snell, and L Wan, “RAF-MEK-ERK pathway in cancer evolution and treatment,” Semin Cancer Biol., vol 85, pp 123–154, Oct 2022, doi: 10.1016/j.semcancer.2021.05.010 [106] H Hua, Q Kong, H Zhang, J Wang, T Luo, and Y Jiang, “Targeting mTOR for cancer therapy,” J Hematol Oncol.J Hematol Oncol, vol 12, no 1, p 71, Dec 2019, doi: 10.1186/s13045-019-0754-1 [107] D A Fruman, H Chiu, B D Hopkins, S Bagrodia, L C Cantley, and R T Abraham, “The PI3K Pathway in Human Disease,” Cell, vol 170, no 4, pp 605–635, Aug 2017, doi: 10.1016/j.cell.2017.07.029 [108] M S Pino and D C Chung, “The Chromosomal Instability Pathway in Colon Cancer,” Gastroenterology, vol 138, no 6, pp 2059–2072, May 2010, doi: 10.1053/j.gastro.2009.12.065 [109] W Zhao et al., “The CDK inhibitor AT7519 inhibits human glioblastoma cell growth by inducing apoptosis, pyroptosis and cell cycle arrest,” Cell Death Dis., vol 14, no 1, p 11, Jan 2023, doi: 10.1038/s41419-022-05528-8 [110] K.-W Lu et al., “Allyl Isothiocyanate (AITC) Induces Apoptotic Cell Death In Vitro and Exhibits Anti-Tumor Activity in a Human Glioblastoma GBM8401/luc2 Model,” Int J Mol Sci., vol 23, no 18, p 10411, Sep 2022, doi: 10.3390/ijms231810411 [111] T.-C Hsia et al., “Bisdemethoxycurcumin Induces Cell Apoptosis and Inhibits Human Brain Glioblastoma GBM 8401/Luc2 Cell Xenograft Tumor in Subcutaneous Nude Mice In Vivo,” Int J Mol Sci., vol 23, no 1, p 538, Jan 2022, doi: 10.3390/ijms23010538 [112] S K Hobbs et al., “Regulation of transport pathways in tumor vessels: Role of tumor type and microenvironment,” Proc Natl Acad Sci., vol 95, no 8, pp 4607– 4612, Apr 1998, doi: 10.1073/pnas.95.8.4607 [113] K Lenting, R Verhaak, M Ter Laan, P Wesseling, and W Leenders, “Glioma: experimental models and reality,” Acta Neuropathol (Berl.), vol 133, no 2, pp 263– 282, Feb 2017, doi: 10.1007/s00401-017-1671-4 [114] F Akter, B Simon, N L De Boer, N Redjal, H Wakimoto, and K Shah, “Preclinical tumor models of primary brain tumors: Challenges and opportunities,” Biochim Biophys Acta BBA - Rev Cancer, vol 1875, no 1, p 188458, Jan 2021, doi: 10.1016/j.bbcan.2020.188458 [115] T Szatmari et al., “Detailed characterization of the mouse glioma 261 tumor model for experimental glioblastoma therapy,” Cancer Sci., vol 97, no 6, pp 546–553, Jun 2006, doi: 10.1111/j.1349-7006.2006.00208.x [116] V Genoud et al., “Responsiveness to anti-PD-1 and anti-CTLA-4 immune checkpoint blockade in SB28 and GL261 mouse glioma models,” OncoImmunology, vol 7, no 12, p e1501137, Dec 2018, doi: 10.1080/2162402X.2018.1501137 [117] V L Jacobs, P A Valdes, W F Hickey, and J A De Leo, “Current Review of in Vivo GBM Rodent Models: Emphasis on the CNS-1 Tumour Model,” ASN Neuro, vol 3, no 3, p AN20110014, Jul 2011, doi: 10.1042/AN20110014 [118] P F Ledur, G R Onzi, H Zong, and G Lenz, “Culture conditions defining glioblastoma cells behavior: what is the impact for novel discoveries?,” Oncotarget, vol 8, no 40, pp 69185–69197, Sep 2017, doi: 10.18632/oncotarget.20193 [119] T M Johanns et al., “Endogenous Neoantigen-Specific CD8 T Cells Identified in Two Glioblastoma Models Using a Cancer Immunogenomics Approach,” Cancer Immunol Res., vol 4, no 12, pp 1007–1015, Dec 2016, doi: 10.1158/23266066.CIR-16-0156 [120] T R Hodges et al., “Mutational burden, immune checkpoint expression, and mismatch repair in glioma: implications for immune checkpoint immunotherapy,” Neuro-Oncol., vol 19, no 8, pp 1047–1057, Aug 2017, doi: 10.1093/neuonc/nox026 [121] R M Samstein et al., “Tumor mutational load predicts survival after immunotherapy across multiple cancer types,” Nat Genet., vol 51, no 2, pp 202– 206, Feb 2019, doi: 10.1038/s41588-018-0312-8 [122] V Letchuman, L Ampie, A H Shah, D A Brown, J D Heiss, and P Chittiboina, “Syngeneic murine glioblastoma models: reactionary immune changes and immunotherapy intervention outcomes,” Neurosurg Focus, vol 52, no 2, p E5, Feb 2022, doi: 10.3171/2021.11.FOCUS21556 [123] S.-M Hede et al., “GFAP promoter driven transgenic expression of PDGFB in the mouse brain leads to glioblastoma in a Trp53 null background,” Glia, vol 57, no 11, pp 1143–1153, Aug 2009, doi: 10.1002/glia.20837 [124] A Richmond and Y Su, “Mouse xenograft models vs GEM models for human cancer therapeutics,” Dis Model Mech., vol 1, no 2–3, pp 78–82, Sep 2008, doi: 10.1242/dmm.000976 [125] F Lin et al., “ABCB1, ABCG2, and PTEN Determine the Response of Glioblastoma to Temozolomide and ABT-888 Therapy,” Clin Cancer Res., vol 20, no 10, pp 2703–2713, May 2014, doi: 10.1158/1078-0432.CCR-14-0084 [126] A T Yeo and A Charest, “Immune Checkpoint Blockade Biology in Mouse Models of Glioblastoma: C HECKPOINT I NHIBITION IN M ODELS OF G LIOMA,” J Cell Biochem., vol 118, no 9, pp 2516–2527, Sep 2017, doi: 10.1002/jcb.25948 [127] M Allen, M Bjerke, H Edlund, S Nelander, and B Westermark, “Origin of the U87MG glioma cell line: Good news and bad news,” Sci Transl Med., vol 8, no 354, Aug 2016, doi: 10.1126/scitranslmed.aaf6853 [128] S Yamada et al., “A method to accurately inject tumor cells into the caudate/putamen nuclei of the mouse brain,” Tokai J Exp Clin Med., vol 29, no 4, pp 167–173, Dec 2004 [129] F Behar, A Ciurea, M Chivu, O Zarnescu, R Radulescu, and D Dragu, “The development of xenograft glioblastoma implants in nude mice brain,” J Med Life, vol 1, no 3, pp 275–286, Aug 2008 [130] M Candolfi et al., “Intracranial glioblastoma models in preclinical neurooncology: neuropathological characterization and tumor progression,” J Neurooncol., vol 85, no 2, pp 133–148, Oct 2007, doi: 10.1007/s11060-007-9400-9 [131] M Vandamme et al., “Response of Human Glioma U87 Xenografted on Mice to Non Thermal Plasma Treatment,” Plasma Med., vol 1, no 1, pp 27–43, 2011, doi: 10.1615/PlasmaMed.v1.i1.30 [132] N Kijima and Y Kanemura, “Mouse Models of Glioblastoma,” Exon Publ., pp 131–139, Sep 2017, doi: 10.15586/codon.glioblastoma.2017.ch7 [133] S Miki et al., “Concomitant administration of radiation with eribulin improves the survival of mice harboring intracerebral glioblastoma,” Cancer Sci., vol 109, no 7, pp 2275–2285, Jul 2018, doi: 10.1111/cas.13637 [134] C Brighi et al., “Comparative study of preclinical mouse models of high-grade glioma for nanomedicine research: the importance of reproducing blood-brain barrier heterogeneity,” Theranostics, vol 10, no 14, pp 6361–6371, 2020, doi: 10.7150/thno.46468 [135] E Yada, S Wada, S Yoshida, and T Sasada, “Use of patient-derived xenograft mouse models in cancer research and treatment,” Future Sci OA, vol 4, no 3, p FSO271, Mar 2018, doi: 10.4155/fsoa-2017-0136 [136] C R Chokshi, N Savage, C Venugopal, and S K Singh, “A Patient-Derived Xenograft Model of Glioblastoma,” STAR Protoc., vol 1, no 3, p 100179, Dec 2020, doi: 10.1016/j.xpro.2020.100179 [137] J Lee et al., “Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than serum-cultured cell lines,” Cancer Cell, vol 9, no 5, pp 391–403, May 2006, doi: 10.1016/j.ccr.2006.03.030 [138] X F Fei et al., “Development of clinically relevant orthotopic xenograft mouse model of metastatic lung cancer and glioblastoma through surgical tumor tissues injection with trocar,” J Exp Clin Cancer Res., vol 29, no 1, p 84, Dec 2010, doi: 10.1186/1756-9966-29-84 [139] M Patrizii, M Bartucci, S R Pine, and H E Sabaawy, “Utility of Glioblastoma Patient-Derived Orthotopic Xenografts in Drug Discovery and Personalized Therapy,” Front Oncol., vol 8, p 23, Feb 2018, doi: 10.3389/fonc.2018.00023 [140] K S Sung et al., “Success of tumorsphere isolation from WHO grade IV gliomas does not correlate with the weight of fresh tumor specimens: an immunohistochemical characterization of tumorsphere differentiation,” Cancer Cell Int., vol 16, no 1, p 75, Dec 2016, doi: 10.1186/s12935-016-0350-1 [141] G V Trunova, O V Makarova, M E Diatroptov, I M Bogdanova, L P Mikchailova, and S O Abdulaeva, “Morphofunctional Characteristic of the Immune System in BALB/c and C57Bl/6 Mice,” Bull Exp Biol Med., vol 151, no 1, pp 99– 102, May 2011, doi: 10.1007/s10517-011-1268-1 [142] A Fukushima et al., “Genetic background determines susceptibility to experimental immune-mediated blepharoconjunctivitis: Comparison of Balb/c and C57BL/6 mice,” Exp Eye Res., vol 82, no 2, pp 210–218, Feb 2006, doi: 10.1016/j.exer.2005.06.010 [143] S.-J Oh et al., “Human U87 glioblastoma cells with stemness features display enhanced sensitivity to natural killer cell cytotoxicity through altered expression of NKG2D ligand,” Cancer Cell Int., vol 17, no 1, p 22, Dec 2017, doi: 10.1186/s12935-017-0397-7 [144] A Karami, M Hossienpour, E Mohammadi Noori, M Rahpyma, K Najafi, and A Kiani, “Synergistic Effect of Gefitinib and Temozolomide on U87MG Glioblastoma Angiogenesis,” Nutr Cancer, vol 74, no 4, pp 1299–1307, Apr 2022, doi: 10.1080/01635581.2021.1952441 [145] T D Pokrovska et al., “External Beam Radiation Therapy and Enadenotucirev: Inhibition of the DDR and Mechanisms of Radiation-Mediated Virus Increase,” Cancers, vol 12, no 4, p 798, Mar 2020, doi: 10.3390/cancers12040798 [146] Z Xu, M Kader, R Sen, and D G Placantonakis, “Orthotopic Patient-Derived Glioblastoma Xenografts in Mice,” in Glioblastoma, D G Placantonakis, Ed., in Methods in Molecular Biology, vol 1741 New York, NY: Springer New York, 2018, pp 183–190 doi: 10.1007/978-1-4939-7659-1_14 [147] A Sadeghipour and P Babaheidarian, “Making Formalin-Fixed, Paraffin Embedded Blocks,” in Biobanking, W H Yong, Ed., in Methods in Molecular Biology, vol 1897 New York, NY: Springer New York, 2019, pp 253–268 doi: 10.1007/978-1-4939-8935-5_22 [148] A H Fischer, K A Jacobson, J Rose, and R Zeller, “Paraffin Embedding Tissue Samples for Sectioning,” Cold Spring Harb Protoc., vol 2008, no 5, p pdb.prot4989, May 2008, doi: 10.1101/pdb.prot4989 [149] “Harris Hematoxylin and Eosin (HE) Staining Protocol.” https://www.ihcworld.com/_protocols/special_stains/HE_Harris.htm (accessed May 19, 2023) [150] “Protocol Online: Xenograft Tumor Model Protocol.” Accessed: May 14, 2023 [Online] Available: http://www.protocol-online.org/prot/Protocols/Xenograft-TumorModel-Protocol-3810.html [151] Mouse Genome Sequencing Consortium, “Initial sequencing and comparative analysis of the mouse genome,” Nature, vol 420, no 6915, pp 520–562, Dec 2002, doi: 10.1038/nature01262 [152] A Olejniczak, M Szaryńska, and Z Kmieć, “In vitro characterization of spheres derived from colorectal cancer cell lines,” Int J Oncol., Nov 2017, doi: 10.3892/ijo.2017.4206 [153] F Maradonna et al., “A zebrafish HCT116 xenograft model to predict anandamide outcomes on colorectal cancer,” Cell Death Dis., vol 13, no 12, p 1069, Dec 2022, doi: 10.1038/s41419-022-05523-z [154] Y Diao, “Inhibition of angiogenesis and HCT-116 xenograft tumor growth in mice by kallistatin,” World J Gastroenterol., vol 13, no 34, p 4615, 2007, doi: 10.3748/wjg.v13.i34.4615 [155] D Wang et al., “Detection of CD133 expression in U87 glioblastoma cells using a novel anti-CD133 monoclonal antibody,” Oncol Lett., vol 9, no 6, pp 2603–2608, Jun 2015, doi: 10.3892/ol.2015.3079 [156] J D Lathia, S C Mack, E E Mulkearns-Hubert, C L L Valentim, and J N Rich, “Cancer stem cells in glioblastoma,” Genes Dev., vol 29, no 12, pp 1203– 1217, Jun 2015, doi: 10.1101/gad.261982.115 [157] M J Clark et al., “U87MG Decoded: The Genomic Sequence of a Cytogenetically Aberrant Human Cancer Cell Line,” PLoS Genet., vol 6, no 1, p e1000832, Jan 2010, doi: 10.1371/journal.pgen.1000832 [158] S Sharma et al., “Human telomerase is directly regulated by non-telomeric TRF2G-quadruplex interaction,” Cell Rep., vol 35, no 7, p 109154, May 2021, doi: 10.1016/j.celrep.2021.109154 [159] V Patil, J Pal, and K Somasundaram, “Elucidating the cancer-specific genetic alteration spectrum of glioblastoma derived cell lines from whole exome and RNA sequencing,” Oncotarget, vol 6, no 41, pp 43452–43471, Dec 2015, doi: 10.18632/oncotarget.6171 [160] A Maeda et al., “The Innate Cellular Immune Response in Xenotransplantation,” Front Immunol., vol 13, p 858604, Mar 2022, doi: 10.3389/fimmu.2022.858604 [161] I A Rota and F Dhalla, “FOXN1 deficient nude severe combined immunodeficiency,” Orphanet J Rare Dis., vol 12, no 1, p 6, Dec 2017, doi: 10.1186/s13023-016-0557-1 [162] M Jivrajani, M V Shaikh, N Shrivastava, and M Nivsarkar, “An improved and versatile immunosuppression protocol for the development of tumor xenograft in mice,” Anticancer Res., vol 34, no 12, pp 7177–7183, Dec 2014 [163] E N Bogdándi et al., “Effects of Low-Dose Radiation on the Immune System of Mice after Total-Body Irradiation,” Radiat Res., vol 174, no 4, pp 480–489, Oct 2010, doi: 10.1667/RR2160.1 [164] C Constantinides, R Mean, and B J Janssen, “Effects of Isoflurane Anesthesia on the Cardiovascular Function of the C57BL/6 Mouse,” ILAR J Natl Res Counc Inst Lab Anim Resour., vol 52, pp e21–e31, 2011 [165] K Hohlbaum, B Bert, S Dietze, R Palme, H Fink, and C Thöne-Reineke, “Impact of repeated anesthesia with ketamine and xylazine on the well-being of C57BL/6JRj mice,” PLOS ONE, vol 13, no 9, p e0203559, Sep 2018, doi: 10.1371/journal.pone.0203559 [166] M Ghandili and S Munakomi, “Neuroanatomy, Putamen,” in StatPearls, Treasure Island (FL): StatPearls Publishing, 2023 Accessed: May 26, 2023 [Online] Available: http://www.ncbi.nlm.nih.gov/books/NBK542170/ [167] F Jin, H J Jin-Lee, and A J Johnson, “Mouse Models of Experimental Glioblastoma,” in Gliomas, Brain Tumor Center of Excellence, Wake Forest Baptist Medical Center Comprehensive Cancer Center, Winston Salem, NC, USA and W Debinski, Eds., Exon Publications, 2021, pp 15–46 doi: 10.36255/exonpublications.gliomas.2021.chapter2 [168] K F Hübner, N Gengozian, and T A Gilman, “Suppression and Recovery of the Immune Response and CFU Potential of Mice during and after Chronic Low-dose-rate Gamma-irradiation,” Int J Radiat Biol Relat Stud Phys Chem Med., vol 28, no 4, pp 301–312, Jan 1975, doi: 10.1080/09553007514551091 [169] F Karimi Dermani, M Saidijam, R Amini, A Mahdavinezhad, K Heydari, and R Najafi, “Resveratrol Inhibits Proliferation, Invasion, and Epithelial–Mesenchymal Transition by Increasing miR‐ 200c Expression in HCT‐ 116 Colorectal Cancer Cells,” J Cell Biochem., vol 118, no 6, pp 1547–1555, Jun 2017, doi: 10.1002/jcb.25816 [170] K Kai et al., “Maintenance of HCT116 colon cancer cell line conforms to a stochastic model but not a cancer stem cell model,” Cancer Sci., vol 100, no 12, pp 2275–2282, Dec 2009, doi: 10.1111/j.1349-7006.2009.01318.x [171] K M Brown, A Xue, A Mittal, J S Samra, R Smith, and T J Hugh, “Patientderived xenograft models of colorectal cancer in pre-clinical research: a systematic review,” Oncotarget, vol 7, no 40, pp 66212–66225, Oct 2016, doi: 10.18632/oncotarget.11184 [172] J Jung, “Human Tumor Xenograft Models for Preclinical Assessment of Anticancer Drug Development,” Toxicol Res., vol 30, no 1, pp 1–5, Mar 2014, doi: 10.5487/TR.2014.30.1.001 [173] J R Maisin, G B Gerber, J Vankerkom, A Wambersie, J R Maisin, and A Wambersie, “Survival and Diseases in C57BL Mice Exposed to X Rays or 3.1 MeV Neutrons at an Age of or 21 Days,” Radiat Res., vol 146, no 4, p 453, Oct 1996, doi: 10.2307/3579307 [174] M S Dorasamy, A Ab, K Nellore, and P.-F Wong, “Synergistic inhibition of melanoma xenografts by Brequinar sodium and Doxorubicin,” Biomed Pharmacother., vol 110, pp 29–36, Feb 2019, doi: 10.1016/j.biopha.2018.11.010 [175] S M Irtenkauf et al., “Optimization of Glioblastoma Mouse Orthotopic Xenograft Models for Translational Research,” Comp Med., vol 67, no 4, pp 300–314, Aug 2017 [176] S A Richard, Y Ye, H Li, L Ma, and C You, “Glioblastoma multiforme subterfuge as acute cerebral hemorrhage: A case report and literature review,” Neurol Int., vol 10, no 1, Apr 2018, doi: 10.4081/ni.2018.7558 [177] F Seker-Polat, N Pinarbasi Degirmenci, I Solaroglu, and T Bagci-Onder, “Tumor Cell Infiltration into the Brain in Glioblastoma: From Mechanisms to Clinical Perspectives,” Cancers, vol 14, no 2, p 443, Jan 2022, doi: 10.3390/cancers14020443 [178] N A De Vries, J H Beijnen, and O Van Tellingen, “High-grade glioma mouse models and their applicability for preclinical testing,” Cancer Treat Rev., vol 35, no 8, pp 714–723, Dec 2009, doi: 10.1016/j.ctrv.2009.08.011 [179] Y Rong, D L Durden, E G Van Meir, and D J Brat, “‘Pseudopalisading’ Necrosis in Glioblastoma: A Familiar Morphologic Feature That Links Vascular Pathology, Hypoxia, and Angiogenesis,” J Neuropathol Exp Neurol., vol 65, no 6, pp 529–539, Jun 2006, doi: 10.1097/00005072-200606000-00001 [180] “Immunohistopathological and neuroimaging characterization of murine orthotopic xenograft models of glioblastoma multiforme recapitulating the most salient features of human disease,” Histol Histopathol., no 24, pp 879–891, Tháng Năm 2009, doi: 10.14670/HH-24.879 [181] D J Brat et al., “Pseudopalisades in Glioblastoma Are Hypoxic, Express Extracellular Matrix Proteases, and Are Formed by an Actively Migrating Cell Population,” Cancer Res., vol 64, no 3, pp 920–927, Feb 2004, doi: 10.1158/00085472.CAN-03-2073 PHỤ LỤC Bảng PL- Thành phần môi trường nuôi cấy DMEM pH = 7.0-7.4 STT Hóa chất Bột mơi trường DMEM NaHCO3 HCl 1M NaOH 1M Nước cất Tổng cộng Khối lượng (g) 13.40 3.70 Vừa đủ Vừa đủ 1000 mL 1000 mL Ghi Đã hấp tiệt khuẩn Lọc qua màng lọc 0.22 μm Bảo quản 4oC Bảng PL- Thành phần mơi trường ni cấy DMEM có bổ sung 10% FBS 1% Penicillin G-Streptomycin STT Hóa chất Thể tích (mL) Dung dịch DMEM pH = 7.0500 7.4 Dung dịch FBS 50 Dung dịch Penicillin G5 Streptomycin Tổng cộng 555 Ghi Bảo quản 4oC Bảng PL- Công thức pha dung dịch PBS STT Hóa chất Viên nén PBS Nước cất Tổng cộng Thành phần 10 viên Vừa đủ 1000 mL 1000 mL Ghi Hấp tiệt khuẩn Bảo quản 4oC Bảng PL- Cơng thức pha lỗng dung dịch Ketamin tiêm chuột STT Hóa chất Tỷ lệ Ghi Ketamin (50mg/10mL) Bảo quản 4oC Nước muối sinh lý pha tiêm 0.9% Tổng cộng Tùy theo số lượng Pha trước sử dụng chuột