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Nghiên ứu xây dựng quy trình hẩn đoán đột biến gene kras

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Tiêu đề Nghiên ứu xây dựng quy trình hẩn đoán đột biến gene kras
Tác giả Ts. Nguyễn Văn B, Pgs. Ts. Trần Văn A
Trường học University of Science
Chuyên ngành Biotechnology
Thể loại Thesis
Năm xuất bản 2013
Thành phố Hanoi
Định dạng
Số trang 76
Dung lượng 3,46 MB

Nội dung

Yoshino, T., et al., TAS-102 monotherapy for pretreated metastatic colorectal cancer: a double-blind, randomised, placebo-controlled phase 2 trial.. Colorectal cancer in inflammatory bowe

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   2 2011 - 2013



 2

 5

 6

 7

 10

 11

 14

 14

 14

 14

 15

 17

 17

 18

 19

 23

 34

1.5 KRAS   37

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   3 2011 - 2013

          

GENE KRAS 40

 40

 42

 43

 43

 43

2.1.2 Hóa ch 43

 44

2.2 45

 45

 46

 46

 48

2.2.5              -Dual Priming-Oligonucleotide) 49

 52

 56

 56

6 56

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           

KRAS 57

 59

 60

 61

 KRAS 62

             64

 67

 68

 69

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

Trung, khoa S

t n

  l

Tôi sinh-Vi -  -

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DAN

APC: adenomatous polyposis coli

AKT: v-akt murine thymoma viral

CDK: cyclin dependent kinases

DPO: Dual priming oligo

DCC: deleted in colorectal carcinoma

DNA: Deoxyribonucleic acid

DNMT: DNA methyltransferase

DTCS: Dye terminating cycle sequencing

ddNTP: dideoxynucleotide triphosphate

EGFR: Epidermal growth factor receptor

EMEA:European Medicines AgenecyESMO: European society for medical oncology

ERBB: erbv- -b1 avian erythroblastic leukemia viral oncogene homolog EDTA: Ethylene diamine tetraacetic acid ERK: extracellular-signal regulated kinase FDA: Food and drug administration FAP: Familial Adenomatous Polyp FRZ: Frizzled

GSK-e-3 GRB: growth factor receptor-bound protein

GEFs: Guanine nucleotide exchange factors

GAPs: GTPase-activating proteins GTP: Guanosine- -triphosphate 5'GDP: Guanosine diphosphate HNPCC: Hereditary non-polyposis colorectal cancer

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   8 2011 - 2013

HER: human epidermal growth factor

receptor

HAT: Histon acetyltransferase

HDAC: Histon deacetylase

HRM: High Resolution Melting

IBD: Inflammatory bowel disease

IGF1R: insulin-like growth-factor-1

receptors

IRS-1: insulin receptor substrate 1

JUN: jun proto-oncogene

KRAS: Ki V- -ras2 Kirsten rat sarcoma 2

viral oncogene homolog

LRP: lipoprotein receptor-related protein

MEK: mitogene-activated protein kinase

MAPK: mitogene activated protein kinase

MMR: Mismatch repair enzymes

MSH: mutS homolog

MeCP2: methyl CpG binding protein 2

MBD: methyl-CpG binding domain

protein

MLH1: mutL homolog 1

HLH1: hemophagocytic lymphohistiocytosis 1 mt: mutant

NST

OD: Optical density p53: protein p53 p21: protein p21 PI3K: Phosphoinositide 3-kinase PDGFR: platelet-derived growth-factor receptor

PIP2: phosphatidylinositol-4, bisphosphate

5-PIP3: trisphosphate

phosphatidylinositol-3,4,5-PDK: phosphoinositide-dependent kinase PTEN: Phosphatase and tensin

homologue PIK3CA Phosphatidylinositol-4,5-: bisphosphate 3-kinase

PBS: Phosphate buffer saline PCR: Polymerase chain reaction pH: Hydrogene power

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   9 2011 - 2013

MSI: Microsatellite instability

SOS: son of sevenless

SHC: Src homology 2

SDS: Sodium dodecyl sulfate

TE: Tris-EDTA

TGF- transforming growth factor beta 

TGFB:transforming growth factor beta

mTOR: mammalian target of rapamycin

RNA: Ribonucleic acid

RTK: receptor tyrosine kinase

SFRPs: Secreted Frizzled-related proteins TGFBR: transforming growth factor beta receptor

TP53: tumor protein p53 UICC: Union for International Cancer Control

UC: Ulerative colitis



WNT: Wingless-type wt: Wild type

µg Microgram

 Microliter



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   11 2011 - 2013



 14

 15

 17

 23

- 25

-34 [10] 27

 29

 32

 37

- dòng kháng EGFR [6] 38

Hình    49

 -G38A 58 .

-G38A Error! Bookmark not defined -G35 61

KRAS-G34 62

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



 ESMO Cetuximab Panitumumab 

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

gene 

 20-3 T

Trang 18

 guyên nhân gây

Escherichia coli , Helicobacter paratuberculosis, Trong 

Trang 19

gene , n gene

APC [1]

Trang 21

Epigene 

 methyl hóa DNA

       ethyl hóa DNA      methyl (-CH3          (DNMT) [28, 29] gene cytosine trong các

         -phosphate-  methyl hóa cygene 

geneCpG (CpG islands[21, 29, 30]

        gene  

gene

gene [30, 31]



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         



                enzyme H, do 

 gene

              enzyme methyl hóa histon,           

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

gene  : APC TGF- TP53 , ,

Gene APC (adenomatous polyposis coli): 

mã hóa protein APC tham

[24] )

[1] 

 y s  -catenin  ubiq  - catenin 

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 -       gene SMAD2 SMAD3, 

SMAD4 [28]gene SMAD2 và gene

       microRNA-34 cromi RN     

 

-         cro     

            [1, 10] Protein p53  microRNA-34 

  s , 1.6

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 [21, 42] 



gene KRAS, BRAF, PIK3CA [4, , 39, 43- ] 21 45

Oncogen KRAS (Kirsten rat sarcoma viral oncogene homolog):  trên NST 1212.1,  RAS

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              

  - KRAS  

         - ho     

- RAS/ RAF/MEK/ERK  phiên                (2) RAS /PI3K/Rac            

 RAL, RalGDS, 

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   30 2011 - 2013

(homodimerisation/EGFR-EGFR)       (heterodimerisation/EGFR-HER2, EGFR-HER3, EGFR-HER4)  

b b t ho t sau m t th i gian ng n [44] Khi       gene KRAS 

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KRAS 

n

i trong gene

[49]

Gene PI K3CA (phosphatidylinositol-4,5-bisphosphate 3-kinase): 

  , Gene PIK3CA thuc h gene    protein heterodimeric, c u trúc phân t hoàn ch nh c   a PIK3CA c k t h p t    ti  u hòa p85 và ti có ho t tính xúc tác p110 [2]

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   32 2011 - 2013

S ho ng cng tín hi u PIK3CA có th 

  u t ng ho c insulin liên k t v i vùng ngo   

 n ng nhRTKK3CA

            

               

              de



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   33 2011 - 2013

tr  c ti p v i PIK3CA kích ho      ng [2, 21, 43, 48 53], PIK3CA c kích ho t s xúc tác ph n ng phosphoryl hóa chuy     i phosphatidylinositol-4,5-bisphosphate (PIP2) thành phosphatidylinositol-3,4,5-trisphosphate (PIP3) PIP3 ti p t c kích ho t s ho   ng c a phosphoinositide-dependent kinase (PDK) và serine-threonine kinase (AKT) AKT s xúc tác th c

hi n ph n ng phosphoryl hóa nhi u lo i protein trong t bào ch t và trong nhân mà       

hong c a chúng s   i ch

c ch quá trình apoptosis c a t    ng tín hi u PIK3CA   c u hòa

âm tính b i s ho   ng c gene áp ch kh i u PTENa       

Protein PTEN xúc tác quá trình chuy n PIP3 thành PIP2 nên làm gi m n   PIP3 và c ch   s ho ng cng tín hi u PIK3CA [2, 21] 

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          





      

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anti-   

             -866 (PIK3CA inhibitor)      mTOR inhibitor)            N

genegene [43, 57]

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   38 2011 - 2013

Nghiên c u d ch t h c cho th y có kho ng 40-45% b        mang gene KRAS   t bi n [12, 13]y, g n m t n a b nh nhân không có tác    

d ng v i các thu  u tr  n có Các th nghi nh cetuximab và panitumumab ch t s có tác d ng kéo dài th i gian s ng và th th     i gian s ng không b nh cho các b nh nhân gene    KRAS  (hình 1 ) i 10 [5, 11]

dòng kháng EGFR [6] B   c kéo dài th i gian s ng thêm n u   

b nh   t bi    u tr b ng TAS-102  

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