Nghiên cứu đa hình gen TNF α 308 g a và TGF β1 509c t ở bệnh nhân UTBMTBG có HBsAg dương tính tt tiếng anh

MINISTRY OF EDUCATION MINISTRY OF DEFENCE AND TRAINING VIETNAM MILITARY MEDICAL UNIVERSITY PHAN THI HIEN LUONG STUDYING POLYMORPHISM OF TNF-α-308 G>A AND TGF-β1-509C>T IN HEPATOCELLULAR CARCINOMA PATIENTS WITH HBsAg POSITIVE Specialty: INTERNAL MEDICINE Code: 72 01 07 THE SUMMARY OF THE MEDICAL DOTORAL THESIS HANOI – 2020 THIS DOCTORAL THESIS WAS COMPLETED AT VIETNAM MILITARY MEDICAL UNIVERSITY Scientific Instructors: A/PROF Nguyen Ba Vuong A/PROF Tran Viet Tu 1st Contradictor: A/PROF Vu Van Khien 2nd Contradictor: PROF Nong Van Hai 3rd Contradictor:A/PROF Hoang Thanh Tuyen The doctoral thesis will be reported to The Grading and Examinations Committee hold at Vietnam Military Medical University at Searching for the dissertation at: National Library Vietname Military Medical University’s library INTRODUCTION Hepatocellular carcinoma (HCC) is one of the common malignant tumors globally with the main cause is chronic hepatitis B virus (HBV) infection This cancer has a poor prognosis with a short survival time, due to limited early diagnosis The host's single nucleotide polymorphism (SNP) plays an important role in determining the body's immune response to HBV infection, affecting HCC formation Research on gene polymorphism helps stratify risk subjects, improve early detection and treatment results Recently, many studies in the world have shown the influence of TNF-α-308 G> A and TGF – β1-509C>T is associated with an increased HCC risk The carriers of A allele of TNF- α - 308 G>A, T alleles of TGF – β1-509 C>T increase the risk of HCC higher than the other carriers of alleles The polymorphisms influence HCC progression in controlling the concentration of the corresponding cytokines In the microbiological environment, these cytokines interact, support and enhance the effects of each other Vietnam is a country, located in the epidemic zone of hepatitis B virus, having a high incidence of HCC with extremely serious consequences Therefore, the thesis "Studying polymorphism of TNF-α-308 G>A and TGF-β1-509C>T in hepatocellular carcinoma patients with HBsAg positive" was conducted with two objectives Determining the frequency of genotype and allele of TNF-α – 308 and TGF-β1-509 in HCC patients with HBsAg (+) Analysing the association of TNF-α-308, TGF-β1-509 polymorphism with risk of HCC and some clinical and subclinical characteristics of HCC patients * New contributions of this doctoral thesis - This is the first scientific study in Vietnam researching on frequency of genotype, allele of TNF-α-308 G>A, TGF- β1-509 C>T in healthy people, chronic hepatitis B and HCC with HBsAg (+) * TNF-α – 308 G>A + The AA genotype only appeared in HCC group, which was 0.98%, the frequency of alleles of A TNF- α - 308 G>A was low in healthy people, 5.4%, which was similar to Asian population + The frequency of allele A in the healthy group, chronic hepatitis B and HCC was 5.4%, 5.83% and 13.2%, respectively There was differences in the frequency of genotypes GG, GA and A allele among HCC groups compared to healthy, chronic hepatitis B groups * TGF–β1- 509 C>T + The frequency of T allele in the healthy group, HBV and HCC was 52.9%; 61.67% and 63.24%, respectively There was difference in the frequency of T allele in HCC and healthy groups - The study showed that A allele, genotype containing allele A of TNF-α -308 increased the risk of HCC compared with G allele and genotype containing allele G Alen T, genotype containing allele T of TGF-β1-509 increased the risk of HCC compared with C allele and genotype contain C allele Two polymorphisms interacted synergistically with each other increasing the risk of HCC * The doctoral thesis arrangement: This thesis contains 113 pages (without references and appendixes): Introduction: 02 pages, Chapter Overview: 31 pages, Chapter 2: Subjects and methods: 17 pages, Chapter Results: 30 pages, Chapter Discussion: 30 pages, Conclusion: 02 pages, Recommendations: 01 page It includes 46 tables, 23 figures, and 125 references (14 Vietnamese references and 111 English references) CHAPTER 1: OVERVIEW 1.1 Overview of hepatocelular carcinoma Hepatocelular carcinoma is the sixth most common cancer among the 10 common cancers worldwide Located in Southeast Asia, high epidemic area, Vietnam has a high incidence of HCC Corresponding to the high prevalence and mortality rates of HCC, the status of HBV infection is also worrisome According to Robert G et al (2011), Vietnam is one of the countries which has the highest HBV infection rate in the world with nearly 12% of people having HBsAg (+), about 10 million people living with chronic HBV In 2018, GLOBOCAN had notified the ASRI (age specific standard index) of HCC in Vietnam for both sexes is 23,2, ranked fourth globally HCC is closely related to HBV infection However, the reality is that not all patients infected with HBV will develop HCC Therefore, both HBV and patients play important role, they interact with each other to cause the final disease 1.2 Single nucleotide polymorphism (SNP) The human genome, largely preserved during evolution, at least 99.5% of genes between any two individuals will be identical, so the difference is only 0.1% - 0.5% This is the key to creating each individual is unique due to different characteristics (shape, inherited disease ) Many factors in the genome contribute to 0.1% of the difference, of which SNP (single-polymorphism) plays an important role SNP is defined as a single base change on a DNA sequence, this change accounts for a proportion of ≥ 1% in the large community and continue to be inherited for the next generation SNP can be located in different regions of the gene, in the promoter region, it plays an important role in controlling the start and level of transcriptional activity of the gene TNF-α – 308 G>A gene TNF-α gene (Tumor necrosis factor - alpha) is located in locus 21.3 of the short arm chromosome (6p21.3) TNF-α gene has many polymorphism, especially the polymorphism at -308 in the promoter region plays an important role This SNP has types of alleles: the allele G, the other rarer type, the allele A The frequency of genotypes and alleles of TNF-α 308G>A polymorphisms varied among different populations TGF-β1- 509C>T gene TGF-β1 gene (Transforming Growth Factor, Beta 1) is located on the long arm of chromosome 19 (19q13.1-13.3) Many single polymorphisms have been identified in the TGF-β1 gene, of which SNP at the C-509T site at the promoter region has important role It is related to gene transcription capability, producing TGFβ plasma The frequency of allele and genotype of TGF-β1-509 is quite diverse in different populations 1.3 TNFα-308G>A, TGF-β1-509C>T and HCC risk Recently, many studies in the world have shown the influence of polymorphism of TNF-α -308 G> A, TGF-β1-509 C>T with the risk of chronic hepatitis B and HCC due to HBV The researchers suggest that these polymorphisms affect the risk of HCC through controlling the concentration of TNFα, TGFβ plasma TNF-α -308 G/A gene Teixeira A.C et al (2013) studied the association of TNF-α -308 G/A polymorphism with HCC risk in Brazil in 120 HCC patients and 202 healthy people The result had shown there was an increased HCC risk in people carriers A allele, GA gene of TNF–α-308 compared to people carriers G allele, GG gene with (OR = 1,82; 95%CI = 1,07-3,08; p = 0.0351), and (OR = 2,51; 95%CI = 1,394,51; p = 0,0031) Tsai J F et al (2017) studied the effect of TNF- α - 308 G>A on HCC risk related to HBV in 200 HCC infected with HBV patients and 200 cirrhosis with HBV infected patients, concluded that GA gene carriers versus GG carriers have an increased risk of HCC with OR = 2,34; 95% CI (1,29 – 4,25); p = 0,004 TGF-β1-509C>T gene Ma J (2015) studied 393 patients with hepatitis C found that the risk of HCC was higher in patients with TT gene than CC gene with OR = 1,820, p = 0.03 Patients carrier T allele compared with C allele increased the risk of HCC with OR= 1,383, p = 0.028 Combination TNFα-308G>A, TGF-β1-509C>T HCC, complex disease, is affected by many factors, of which the role of a gene is very important, but in fact, no single polymorphism can fully explain the complex mechanisms of disease, each SNP plays only a very small role in the development of disease Therefore, the new trend is studying the combination of genotypes and alleles of different SNPs with the risk of disease, help stratify This helps increase the value of early detection and improve the effectiveness of disease treatment Bei C.H (2014) evaluated the effect of SNP include IL-2, IFN-γ, IL-1β, IL-6 and IL10 genes with risk of HCC in China in 720 HCC patients and 784 healthy people, had concluded that although not a single SNP increased the risk of HCC, but combination of SNP had increased HCC risk with OR = 1,821; 95%CI = 1,078 3,075 The authors said that interactions between SNPs increase the risk of HCC El Din N.G (2017) assessed the effects of TNF-α -308 and TGF –β1-509 on cirrhosis progression due to HCV showed that combination of TNF-α- 308 AA and TGF-β1-509 TT compared with TNF (GG) + TGF (CC) increases risk of cirrhosis with OR = 6,4; 95%CI = 1,490 – 27,641; p = 0,013 The results showed that these was a synergistic effect of genes because TNF-α alone increased the risk of cirrhosis by 2.8 times, TGF-β1 increased the risk of cirrhosis by 2.9 times 1.4 Studies TNFα-308G>A and TGF-β1-509C>T in Vietnam Recently, the role of polymorphisms for HCC has begun to be studied in Vietnam However, as far as we know, there have been no studies on TNF- α- 308 and TGF-β- 509 in HCC patients In 2012, Dunstan studied in 2350 healthy Vietnamese people, genetic analysis was conducted in England The result showed that the percentage of A allele TNF-α - 308 was 7% Tran Dinh Tri (2017) studied the gene TNF-α - 308 G>A in patients with stomach cancer showed that the percentage of allele A was 17.1% Such research results have been noted that the percentage of A allele of TNF-α - 308 in the Vietnamese population is low, similar to other Asian countries The polymorphism TGF-β1- 509 C>T has been extensively studied in the world with certain roles in many diseases However, in the Vietnamese population, although we have tried to search, no studies on this polymorphism have been published CHAPTER 2: SUBJECTS AND METHODS 2.1 Subjects The study included 102 HCC infected with HBV patients, 60 chronic hepatitis B patients, 102 healthy people 2.1.1 Study group Inclusion criteria: - Diagnosis of HCC was base on the guidance of the Ministry of Health of Vietnam in 2012, has of the following criteria + Anatomical or histopathological evidence of histology + Typical image on contrast CT scan or contrast MRI with αFP ≥ 400 ng/ml - Patients with HBsAg (+) - The patient was first diagnosed with HCC Exclusion criteria - Patients with HIV infection - Patients with HCV infection - Patients with alcoholic liver disease The liver is damaged by drugs or chemicals Metastatic liver cancer from other organs Patients were need liver biopsy for diagnosis but having contraindication to liver biopsy (platelets A polymorphism 3.2.1 Frequency of genotypes, alleles of TNF-α-308 G> A - The frequency of GG genotype has the lowest rate of 75.51% in the HCC group, the highest rate in the healthy group is 89.22% - Alen A has the highest rate in the HCC group 13.24%, lower in the hepatitis B group 5.83% and lowest in the healthy group 5.4% - There was a significant difference in the frequency of GG, GA genotypes and alleles in HCC group compared to healthy group 3.3.1 TNF-α -308 polymorphism and HCC 3.3.1.1 TNF-α -308 polymorphism and HCC risk The control group is healthy people Table 3.13 TNF-α-308 polymorphsim and HCC risk ( Control is healthy people) Genotype and allele HCC n=102 GG GA AA GA + AA 76 25 26 Healthy OR (95%CI) n=102 Genotype (n) 91 Ref 11 2,721 (1,258 – 5,888) 11 2,830 (1,313 – 6,100) Allele (2n) p 0,009 0,006 12 Alen G Alen A 177 27 193 11 Ref 2,676 (1,290 – 5,555) 0,006 ( Ref- Reference) - The GA genotype, the (GA + AA) genotype compared to the GG genotype, increases HCC risk with OR = 2,721; 95% CI (1,258 – 5,888); p = 0,009 and OR = 2,830; 95% CI (1,313 –6,100); p = 0,006 - A allele was significantly associated with increased risk HCC, compared to G allele, OR= 2,676; 95% CI (1,290 – 5,555); p = 0,006 The control group is chronic hepatits B + healthy people Table 3.14 TNF-α-308G>A and HCC risk ( Control is chronic hepatitis B + healthy ) Genotype and allele HCC n= 102 GG GA AA AA + AG 76 25 26 Alen G Alen A 177 27 Without HCC n = 162 Genotype (n) 144 18 18 Allele (2n) 306 18 OR (95%CI) p Ref 2,632 (1,351 – 5,125) 2,737 (1,412 – 5,306) 0,004 0,002 Ref 2,593 (1,389 – 4,842) 0,002 - GA genotype, (GA + AA) genotype compared to GG genotype, increases HCC risk, with OR = 2,632; 95% CI (1,351 – 5,125); p = 0,004 and OR = 2,737; 95% CI (1,412 – 5,306); p = 0,002 - A allele was significantly associated with increased risk HCC, compared to G allele, OR =2,593; 95% CI (1,389 – 4,842); p = 0,002 The control group is chronic hepatits B Table 3.15 TNF-α-308G>A polymorphism and HCC risk (Control is chronic hepatitis B patients) Genotype and allele HCC n= 102 GG GA AA 76 25 Chronic hepatitis OR (95%CI) B (n= 60) Genotype (n) 53 Ref 2,491 (1,004 – 6,178) - p 0,044 - 13 AA + AG 26 Alen G Alen A 177 27 2,590 (1,048 – 6,405) Allele (2n) 113 Ref 2,462 (1,038 – 5,843) 0,035 0,036 - The GA genotype, the (GA + AA) genotype compared to the GG genotype, increases the risk of HCC, OR = 2,491; 95% CI (1,004 – 6,178); p = 0,044; OR = 2,590; 95% CI (1,048 – 6,405); p = 0,035 - A allele was significantly associated with increased risk HCC, compared to G allele, OR = 2,462; 95% CI (1,038-5,843); p = 0,036 3.2.1.2 TNF-α-308 G>A polymorphism and clinical, subclinical, and stage of HCC Tables 3.16, 3.17, 3.18, 3.19 and 3.20 showed that there was no difference between TNF-α-308G>A polymorrphism with some clinical and subclinical characteristics such as age, tumor characteristics, platelets, αFP, HBV DNA, portal venous thrombosis, metastasis, HCC stage 4.3 TGF-β1-509 C>T polymorphism and HCC risk 4.3.1 Frequency of genotypes, alleles of TGF-β1-509 C>T - CT genotype predominates in all groups T allele in HCC patients (63,24%), chronic hepatitis B (61,67%), healthy people (52,94%) - There was a significant difference in the frequency of alleles in HCC group compared to healthy group, with p = 0.035 4.3.2 TGF-β1-509 C>T polymorphism and HCC 4.3.2.1 TGF-β1-509 C>T polymorphism and HCC risk The control group is healthy people Table 3.21 TGF-β1-509 C> T polymorphism and HCC risk (Control is healthy people) Genotype and allele CC CT TT HCC n= 102 14 47 41 Healthy OR (95%CI) n=102 Genotype (n) 23 Ref 50 1,544 (0,712 – 3,351) 29 2,323 (1,026 – 5,258) p 0,270 0,041 14 CC TT + CT CT + CC TT 14 88 61 41 Alen C Alen T 75 129 23 Ref 79 1,830 (0,881 – 3,799) 73 Ref 29 1,692 (0,943 – 3,036) Allele (2n) 96 Ref 108 1,529 (1,029 – 2,271) 0,102 0,077 0,035 - The TT genotype compared to the CC genotype, increases the risk of HCC, with OR = 2,323; 95% CI (1,026 – 5,258); p = 0,041 - T allele was significantly associated with increased risk HCC, compared to C allele, OR = 1,529; 95% CI (1,029 – 2,271); p = 0,035 Tables 3.22 and 3.23 genotypes and alleles of TGF-β1-509 did not affect the risk of HCC, when using control groups were without HCC, chronic hepatitis B 4.3.2.2 TGF-β1-509 C>T polymorphism and clinical, subclinical, and stage of HCC Tables 3.24, 3.25, 3.26, 3.27 and 3.28 showed that there was no difference between TGF-β1-509C>T polymorphism with some clinical and subclinical characteristics such as age, tumor characteristics, platelets, αFP, HBV DNA, portal venous thrombosis, metastasis, HCC stage 4.4 Combination of TNF-α-308 G>A, TGF-β1-509 C>T polymorphisms We see GG genotype of TNF-α-308 G>A and CC genotype of TGF-β1-509 C>T appeared significantly more in the without HCC group than the HCC group The genotype containing allele A of TNFα - 308 and genotype containing allele T of TGF-β1-509 increased the risk of HCC compared to the remaining genotypes Therefore, we consider GG to be a good genotype compared to AA and CC as a good genotype compared to TT, we divide the appearance of two genes as follows: - Good combination: having genes GG and CC are homozygous - Moderate combination: having gene GG or CC in homozygous 15 - Bad combination: neither GG gene nor CC gene in homozygous 4.4.1 TNF-α–308G>A, TGF-β1-509C>T polymorphism and HCC risk The control group is healthy people Table 3.32 TNF-α – 308 G>A, TGF-β1-509 C> T polymorphism and HCC risk (Control is healthy people) Genotype and allele Good combination Moderate combination Bad combination HCC n= 102 72 21 Healthy n=102 23 68 11 OR (95%CI) p Ref 2,706 (1,169 – 6,261) 4,879 (1,688 – 14,098) 0,017 0,003 The control group is healthy people - Moderate combination, bad combination increases the HCC risk compared to good combination, OR=2,706; 95%CI (1,169 – 6,261);p=0,017 and OR=4,879; 95%CI (1,688 – 14,098); p= 0,003 The control group is chronic hepatits B + healthy people Table 3.33 TNF-α – 308 G>A, TGF-β1-509 C> T polymorphism and HCC risk (Control is healthy people + healthy people) Genotype and allele HCC n= 102 Good combination Moderate combination Bad combination 72 21 Without HCC n = 162 32 112 18 OR (95%CI) p Ref 2,286 (1,031 – 5,070) 4,148 (1,571– 10,956) 0,038 0,003 - The moderate, bad combination increases HCC risk compared to good combination, with OR = 2,286; 95% CI (1,031 – 5,070); p = 0,038 and OR = 4,148; 95% CI (1,571 – 10,956); p = 0,003 When using control group was chronic hepatitis B patient, we did not see an association for HCC risk with polymorphisms 4.4.2 The relation between combination of polymorphisms and some clinical, subclinical characteristics of HCC patients According to tables 3.35, 3.36, 3.37, 3.38, and 3.39, there was no effect of combining polymorphisms with some clinical, 16 subclinical characteristics: age, tumor characteristics, platelets, αFP, HBV DNA, thrombosis portal vein, metastases, HCC stage CHAPTER 4: DISCUSSION 4.1 Clinical and subclinical characteristics of HCC patients HCC patients had an average age of 57.4 ± 9.7 The youngest is 37 years old, the oldest is 80 years old The majority of patients aged 51-70 years old were 67.2% Male / female ratio: 11.8 /1 At the time of diagnosis, the percentage of patient without symptoms was 13,7% The average size of total tumor: 10,80 ± 12,02 cm, size tumor 10 cm was high percentage (37.3%), portal vein thrombosis (22.5%), other metastases (19.6%) Subclinical indicators fluctuate, αFP < 400 ng/ml (52.9%), HBV-DNA ≥ 104 (66.7%) The majority of patients were in stage Okuda II (58.8%), Barcelona B (35.2%), Barcelona C (36.3%) The rate of patients detected at an early stage was low rate such as Okuda I (36.3%), Barcelona A (27.5%) As we know chronic HBV infection is the most important risk factor for HCC, our study subjects were HCC patients with HBsAg (+), but at the time of diagnosis only 49% HCC patients knew themselves have been infected with HBV before The above analysis reflects objectively and accurately the situation of HCC management in Vietnam is still weak This result is due to people's ignorance and the lack of necessary screening programs for high-risk subjects such as people infected HBV Our conclusions are similar to Robert G (2012), when studying the situation of liver disease in Vietnam The above results show that the majority of patients with HCC are diagnosed late This is indeed a barrier to treatment, leading to poor prognosis 4.2 TNF-α-308 G>A polymorphism 17 4.2.1 The frequency of allele and genotypes of TNF-α-308 G>A, polymorphisms - The frequency of GG genotypes was dominant in all groups GG genotype was highest in healthy groups, GA was highest in HCC There was a difference in the frequency of GG and GA genotypes between HCC group with healthy people and chronic hepatitis B (p T polymorphisms - The CT genotype predominates in all groups There was no difference when comparing the frequency of CC, CT, TT genotypes in research groups - The frequency of allele T in the healthy group, chronic hepatitis B and HCC was 52.94%; 61.67%; 63.24% There was a difference in frequency of T allele between HCC and healthy groups, with p T polymorphism and HCC risk The control group is healthy people: - TT genotype has a higher risk of HCC than CC genotype, with OR = 2,323; 95% CI (1,026 – 5,258); p = 0,041 - People with T allele have a higher risk of HCC than those with C allele, with OR = 1,529; 95% CI (1,029 – 2,271); p = 0,035 Some studies have found that TT and T alleles are associated with an increase in TGF-β levels and increased risk of HCC like Radwan M.I (2012), Ma J (2015) However, according to Qi P (2009) found that the new CC and C allele played this role Guo Y et al (2013) conducted a meta-analysis based on studies including 2030 HCC patients and 3416 controls, who found TT and T alleles genotypes increased the risk of HCC TT versus CC genotype increases the risk of HCC with OR = 1,61, 95% CI= 1,21 – 2,14, p= 0,0001 CT genotype increases the risk of HCC when compared to CC genotype with OR=1,28, 95% CI = 1,09 – 1,50, p = 0,003 TT + CT increases risk of HCC compared with CC genotype, with OR = 1,4, 95% CI = 1,15 – 1,70, p= 0,0007 This is a valuable evidence to recognize the role of TGF-β1-509 polymorphism in HCC risk In the process of forming HCC from chronic hepatitis, cirrhosis, TGFβ1 is considered as a direct damage agent Experimental studys disrupted the synthesis of TGFβ1 and/or TGFβ signaling pathways had a significant effect on reducing fibrosis Fatlleti E (2008) investigated the effect of TGF-β1 gene and the risk of liver disease, showed that there was difference in the proportion of genotypes of TGF-β1 polymorphism among patients with end-stage liver disease and normal subjects The author noted that the T allele of TGF-β1 causes increased TGFβ production in patients with cirrhosis The study suggested that the polymorphism of TGFβ gene, majority polymorphisms in promoter region, are involved in cirrhosis Lu W.Q (2016) conducted a meta-analysis of 2809 HCC patients and 4802 without HCC, the subgroup analysis applied to studies, 21 which selected patients in the general population showed that TT genotype compared with CC gene increased risk of HCC (OR, 95%CI, p: 1,74, 1,08-2,8, 0,023) Genes (TT + TC) compared with CC increased the risk of HCC (OR, 95%CI, p: 1,48, 1,01-2,17, 0,047) T alleles versus C alleles increase HCC risk (OR, 95%CI, p: 1,35, 1,05-1,74, 0,021) 4.4 Combination of TNF-α-308 G>A, TGF-β1-509 C>T 4.4.1 Relationship between allele and genetype of TNF-α-308 G> A, TGF-β1-509 C> T with HCC risk The control group is healthy people - Bad combination, moderate combination increase HCC risk compared to good combination, OR=4,879; 95%CI (1,688 – 14,098); p = 0,003 and OR = 2,706; 95%CI (1,169 - 6,261); p = 0,017 The control group is chronic hepatits B + healthy people - Bad combination, moderate combination increase HCC risk compared to good combination, OR=4,148; 95%CI (1,571 – 10,956); p = 0,003 and OR = 2,286; 95%CI (1,031 – 5,070); p = 0,038 The result showed that there was an enhanced interaction effect when combining genes with each individual gene as bad genotypes combining increase HCC risk to good genotypes combination (control group wass healthy), OR = 4,879; 95%CI (1,688 – 14,098); p = 0,003 The combined OR was much greater than the risk calculated separately for each polymorphism, such as TNF-α alone increased the risk of HCC by 2.83 times, TGF-β1 increased the risk of HCC by 2,323 times This consistent was summarized by Liongue C (2015) found that TGFβ acted synergistically with TNFα to stimulate fibrous proliferation, promoted the process of ECM (epithelial transition to mesenchyma), and these cytokines interacted mutually to promote HCC formation and metastasis In addition, El Din N.G (2017) concluded that these two polymorphisms had synergistic effects on cirrhosis progression 22 4.4.2 Relationship between TNF-α - 308, TGF-β1-509 with some clinical, subclinical characteristics of HCC patients Although there was a relationship between each polymorphism and combination of TNF-α-308 G>A and TGF-β1-509 C>T with HCC risk in different degrees, we did not detect a significant difference between each polymorphism and their coordination with some clinical and subclinical symptoms such as tumor characteristics (number, differentiation, size), platelets, αFP, HBV- DNA, portal vein thrombosis, metastases, and HCC stage In fact, HCC is a complex disease with many factors, although results from many studies showed the important role of genetic factors in HCC risk However, some polymorphisms are too small to affect clinical disease Indeed, there have been many studies showing that other polymorphisms of TNFα, TGFβ, or many other genes also influenced the risk of developing HCC such as TP53, MDM2, IFNAR1 gene In addition, tumor characteristics, αFP, HBV DNA are greatly influenced by moment of diagnosis (soon or late) and the status of previous treatment HCC is highly malignant, silently evolved, and its growth rate is high, while genetic polymorphism is a more stable genetic element, so it is difficult to have a significant relationship between them is reasonable With the same research model for evaluation the influence of TNF-α-308G>A and TGF-β1509C>T polymorphisms with characteristics of liver tumor, metastasis Radwan M.I (2012) had the same opinion with us This is the first study in Vietnam evaluated the effects of TNFα -308 and TGF-β1-509 with risk of HCC Results showed that allele A, genotype containing A of TNFα - 308 and T alleles, genotype containing T of TGF-β1-509 gene were associated with an increased risk of HCC In particular, the two polymorphisms had synergistic interaction, increased the risk of HCC However, we had not selected control groups had the same age with disease group, all HCC patients were selected in one hospital Therefore, the results should 23 be confirmed in the following studies with the same age in groups and the subjects from many hospitals CONCLUSIONS Studying 102 HCC patients with HBsAg (+), 60 chronic hepatitis B patients and 102 healthy people, we concluded The frequency of allele and genotypes of TNF-α-308 G>A, TGF-β1- 509C>T polymorphisms 1.1 TNF-α – 308 G>A - GG genotype predominates in all groups, highest in healthy group, GA genotype is highest in HCC group There was a difference in the frequency of GG and GA genotypes between HCC and healthy groups, chronic hepatitis B (p T with HCC 2.1 Relationship between allele and genetype of TNF-α-308 G> A, TGF-β1-509 C> T with HCC risk - Allele A and genotypes containing allele A of TNF-α -308 both increase the risk of HCC compared to G allele and genotype containing allele G, when using control group who are healthy people, without HCC people and chronic hepatitis B patients, with 24 OR= 2,46; 95% CI(1,038-5,843); p = 0,036 to OR= 2,83; CI(1,313 – 6,100); p = 0,006 - Allele T and genotypes containing allele T of TGF-β1-509 increase the risk of HCC compared to C allele and genotypes containg allele C, when using control group who are healthy people, with OR = 1,529; 95% CI(1,029 – 2,271); p = 0,035 to OR = 2,323; CI (1,026 – 5,258); p = 0,041 - Two polymorphisms interacted synergistically to increase HCC risk Moderately and bad combinations increased the risk of HCC compared to the good combination when using control groups who were healthy and without HCC, with OR = 2,286; 95%CI (1,031 – 5,070); p = 0,038 to OR= 4,879; 95%CI(1,688 – 14,098); p = 0,003 2.2 Relationship between TNF-α - 308, TGF-β1-509 with some clinical, subclinical characteristics of HCC patients - There was no relationship between each polymorphism and the combination of TNF-α-308G>A and TGF-β1-509C>T with some clinical, subclinical characteristics and HCC stages RECOMMENDATIONS Studying TNF-α-308G>A, TGF-β1-509C>T polymorphisms with the risk of HCC in the prospective study Assessing the influence of TNF-α-308 G>A, TGF-β1-509C> T interacting with environmental risk factors such as living habits, associated other diseases for over process of generating and developing HCC THE INDEX OF DOCTORAL THESIS-RELATED JOURNALS PUBLISHED Phan Thi Hien Luong, Nguyen Ba Vuong, Tran Viet Tu, Luong Thi Lan Anh (2019) The effect of TNF-α-308G>A polymorphism alters the risk of hepatocellular carcinoma, Journal of Military Pharco-Medicine 44(5): 170-176 Nguyen Ba Vuong, Phan Thi Hien Luong, Tran Viet Tu, Luong Thi Lan Anh (2019) The correlation between TGF-β-509 gene polymorphism and risk of hepatocellular carcinoma Journal of Military Pharco-Medicine, 44(5): 51-57 Phan Thi Hien Luong, Luong Thi Lan Anh, Nguyen Hai Linh, Hoang Ha (2019) Using sequensing method to determine of TNF-α-308G/A gene and TGF-β-509 C/T gene polymorphisms in hepatocellular carcinoma Vietnam Medical Journal, 483(topic): 131-139 ... - 308: frequency of allele G, A, gene: AA, AG, GG - TGF- β1- 509: frequency of allele C, T, gene: CC, CT, TT - Two associated genes: frequency of allele (GC, GT, AC, AT); genes: (GGCC, GGTT, GGCT,... hepatitis so that factors that control inflammatory processes such as TNF- α greatly influence these activities Although TNF? ? is a multifunctional cytokine, have inflammatory effect, against viral... A, TGF- β1- 509C> T with HCC 2.1 Relationship between allele and genetype of TNF- α- 308 G> A, TGF- β1- 509 C> T with HCC risk - Allele A and genotypes containing allele A