Studying polymorphism of tnf - Α - 308 g>a and tgf - β1 - 509 c> t in hepatocellular carcinoma patients with hbsag positive

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 6 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 6 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 2 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 1 of the following 2 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 <80 G/l, PT <60%) 2.1.2. Control group 2.1.2.1.Group 1: 60 chronic hepatitis B patients Inclusion criteria: The patients was diagnosed of chronic hepatitis B according to guidance of the Ministry of Health of Vietnam in 2014 HBsAg (+) ≥ 6 months or HBsAg (+) and Anti HBc IgG (+) ALT, AST increase (intermittent or continuous ≥ 6 months) Evidence of progressive histopathological damage (determining by liver biopsy, or liver elasticity measurement or Fibrotest) Exclusion criteria Patients with alcoholic liver disease Patients with HCV infection Patients with HIV infection Patients with contraindication to liver biopsy (platelets <80 G/l, PT <60%). Group 2: 102 healthy people - Blood donors have HBsAg (-), Anti HCV (-), Anti HIV (-) 2.2. RESEARCH METHODS 2.2.1. Study Design: Descriptive cross-sectional study. n = Z21-α/2 2.2.2. Sampling method: Sampling Size Formula: According to Hu Q. (2014), the frequency of allele A TNF-α - 308 in the disease group was 10.2%, the control group was 7.5%. Instead of the formula, the minimum number of patients needed was 97, this study was conducted on 102 patients (n = 102). 2.2.4. Steps to conduct research 2.2.4.1. Collection HCC patients and control groups Subjects were clinically, subclinically examined (hematology, biochemistry, immunology), according to medical record - Abdominal ultrasound: assessment of liver parenchyma (chronic hepatitis B patients) - CT or MRI: tumor characteristics, portal vein thrombosis, metastasis (HCC patients) - Biopsy and histopathological diagnosis + HCC: for atypical tumors was HCC. + Chronic hepatitis B: according to Metavir, select patients with F0-F3 results. Healthy people: 102 blood donors at 103 Military Hospital 2.2.4.3. Analysis TNF-α - 308 and TGF-β1 -509 polymorphism Sequencing 2 genes directly by ABI PRISM 3500 system. The sequence of reference for 2 genes was taken from Genbank database 2.2.5. Research parameters 2.2.5.1. Characteristics of the research group. - Age: agroup ≤ 40, 41-50, 51-60, 61-70 ,> 70 years; average age. - Gender: male and female; male/female ratio. - Symptoms: clinical, subclinical, stage of disease - Imaging diagnosis: liver tumor (location, number, morphology, size) portal vein thrombosis, metastases - Histopathological characteristics: HCC differentiation (high, medium, low). 2.2.5.2. Frequency of allele and genotype of TNF- α - 308 G>A, TGF- β1 – 509 C>T . - TNF-α - 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, GACC, GATT, GACT, AACC, AATT, AACT). 2.2.5.3. Correlation between TNF-α-308, TGF-β1-509 with HCC risk and clinical, subclinical characteristics of HCC patients - Compare the frequency of genotypes and alleles TNF-α - 308 G> A, TGF- β1 - 509 C> T among research groups. - Calculating the risk of HCC in different genotypes and alleles of TNF-α - 308 G> A, TGF- β1-509 C> T, combining 2 genes. We classify good genes with protective effects, bad genes with increasing the risk of HCC: + Genotypic combination type: good combination - both good genes in the homozygous form, medium combination - one good gene in the homozygous form, bad combination - no good gene in the homozygous form. - The relationship between genotypes of each polymorphism, combining two polymorphisms with clinical symptoms, subclinical and HCC stage. 2.2.6. Data processing: the collected data were processed according to the medical statistics using SPSS 20.0 statistical software 2.3. TIME AND LOCATION OF RESEARCH. Research time: October 2016-October 2017. Research locations: Bach Mai Hospital, Hanoi Medical University, Military Medical Hospital 103. 2.4. RESEARCH ETHICS The subjects were fully explained and were willing to participate in the research. The implementation processes were trictly complied with the regulation of the Ministry of Health of Vietnam. Diagram of the study HCC patients with HBsAg (+) (n = 102) Clinical examination, biochemical tests, hematology, microbiology, imaging, histopathology Analysis TNF-α - 308 and TGF-β1 -509 polymorphism by direct sequencing method chronic hepatitis B patients (n = 60) Healthy people (n = 102) Determination of genotype and allele frequency Analysis of genotypic and allele relationship of TNF-α - 308, TGF-β1 -509, combining 2 polymorphisms with HCC risk and clinical and subclinical symptoms of HCC patients CHAPTER 3: RESULTS 3.1. Clinical and subclinical characteristics of HCC patients 3.1.1. Characteristics of age and gender 82.2% of the patients were male, the male/female ratio was 11.8/1. The average age of HCC patients was 57.4 ± 9.7, the most common age was 51-70 was 67.6%. 3.1.2. Clinical and subclinical characteristics At the time of diagnosis, the number of asymptomatic patients (13.7%), right upper quadrant pain (65.7%), fatigue (35.3%), 49% of patients knew they had been infected with HBV before. Most liver tumors in the right lobe (74,5,%), 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%). 3.2. TNF-α-308 G>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 Healthy n=102 OR (95%CI) p Genotype (n) GG 76 91 Ref GA 25 11 2,721 (1,258 – 5,888) 0,009 AA 1 0 - - GA + AA 26 11 2,830 (1,313 – 6,100) 0,006 Allele (2n) Alen G 177 193 Ref Alen A 27 11 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 Without HCC n = 162 OR (95%CI) p Genotype (n) GG 76 144 Ref GA 25 18 2,632 (1,351 – 5,125) 0,004 AA 1 0 - - AA + AG 26 18 2,737 (1,412 – 5,306) 0,002 Allele (2n) Alen G 177 306 Ref Alen A 27 18 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 Chronic hepatitis B (n= 60) OR (95%CI) p Genotype (n) GG 76 53 Ref GA 25 7 2,491 (1,004 – 6,178) 0,044 AA 1 0 - - AA + AG 26 7 2,590 (1,048 – 6,405) 0,035 Allele (2n) Alen G 177 113 Ref Alen A 27 7 2,462 (1,038 – 5,843) 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 HCC n= 102 Healthy n=102 OR (95%CI) p Genotype (n) CC 14 23 Ref CT 47 50 1,544 (0,712 – 3,351) 0,270 TT 41 29 2,323 (1,026 – 5,258) 0,041 CC 14 23 Ref TT + CT 88 79 1,830 (0,881 – 3,799) 0,102 CT + CC 61 73 Ref TT 41 29 1,692 (0,943 – 3,036) 0,077 Allele (2n) Alen C 75 96 Ref Alen T 129 108 1,529 (1,029 – 2,271) 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 TNF-α - 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 2 genes GG and CC are homozygous - Moderate combination: having 1 gene GG or CC in homozygous - 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 HCC n= 102 Healthy n=102 OR (95%CI) p Good combination 9 23 Ref Moderate combination 72 68 2,706 (1,169 – 6,261) 0,017 Bad combination 21 11 4,879 (1,688 – 14,098) 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 Without HCC n = 162 OR (95%CI) p Good combination 9 32 Ref Moderate combination 72 112 2,286 (1,031 – 5,070) 0,038 Bad combination 21 18 4,148 (1,571– 10,956) 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 2 polymorphisms with some clinical, 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 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 <0.05). AA genotype only appeared in the HCC group with the low frequency was 0.98%. The frequency of allele A in the healthy group, chronic hepatitis B and HCC were 5.4%, 5.83% and 13.2%, respectively. There was a difference in the frequency of allele among the HCC groups compared to healthy people and chronic hepatitis B (p <0.05). Dustan S. J. et al (2012) studied 2350 healthy Vietnamese people, found the frequency of allele A was 7%. There was a slight difference between our results. It may be explained that Dustan selected healthy people in South Vietnam while we chose healthy people in North Vietnam. In addition, Dunstan randomly selected healthy people, excluding cases who were infected HBV, HCV and HIV virus. Furthermore, Li Z. (2017) conducted a meta-analysis of 7254 without HCC people and found that the frequency of allele A in healthy Asians was 6.44%, quite similar to our rate was 5.4%. Our results were similar to Radwan’ study (2012). He reseached in 160 healthy people, 152 cirhosis with HCV patients and 128 HCC patients in Egypt and found: the highest frequency of allele A in HCC group was 19.5%, cirrhosis was 15.8%, and healthy was 7.5%. The frequency of GG genotype in healthy group was 85%, cirrhosis was 71.1% and HCC was 64.1%. The AG + AA genotype was lowest in healthy group (15%), higher in cirrhotic group (28.9%) and HCC group (35.9%). The frequency of genotypes, alleles among groups achieved statistical significance with p <0.001 4.2.1. TNF-α-308 G>A polymorphism and HCC risk The control group is healthy people: - The GA genotype compared to the GG genotype, increases HCC risk, with OR = 2,721; 95% CI (1,258 – 5,888); p = 0,009. The GA + AA genotype compared to the GG genotype, increases HCC risk, with OR = 2,830; 95% CI (1,313 – 6,100); p = 0,006. - People carriers of A allele are at higher risk for HCC than those with G alleles, OR= 2,676; 95% CI (1,290 – 5,555); p = 0,006. The control group is chronic hepatits B + healthy people: The GA genotype compared to the GG genotype, increases HCC risk, with OR = 2,632; 95% CI (1,351 – 5,125); p = 0,004. the (GA + AA) genotype compared to the GG genotype, increases HCC risk with 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 with OR =2,593; 95% CI (1,389 – 4,842); p = 0,002. Our finding were similar to Feng.H’ study (2014). A reseach in China with 753 HCC patients and 760 without HCC showed that AA genotype was associated with increased HCC risk, compared to GG genotype, with OR= 5,12; 95% CI = 2,31 – 7,82. GA + AA genotype showed 5,59 fold increased HCC risk. A allele was associated with increased risk HCC, compared to G allele with OR= 4,18; 95% CI = 1,76- 6,97. The authors concluded that TNF-α - 308 was significantly associated with HCC risk in Han polulation. The control group is chronic hepatits B patients - The GA genotype compared to the GG genotype, increases HCC risk, with OR = 2,491; 95% CI (1,004 – 6,178); p = 0,044. The GA + AA genotype compared to the GG genotype, increases HCC risk, with OR = 2,590; 95% CI (1,048 – 6,405); p = 0,035 People carriers of A allele are at higher risk for HCC than those with G alleles, OR = 2,462; 95% CI (1,038-5,843); p = 0,036 Tsai J.F. et al (2017) studied in Taiwan on 200 HCC patients infected with HBV and 200 patients with HBV cirrhosis to assess the effect of TNF- α - 308 and LT-α +252 with HCC risk related to HBV. AG genes increase cancer risk compared with GG gene (OR = 2,85, 95% CI = 1,60-5,6, p = 0,004). The author also claimed that HBV was a carcinogenic virus, which itself stimulated TNFα production. Thus, when people were infected chronic HBV, both the host and the virus contribute to increased TNF-α production, thereby increasing the risk of HCC. Xiao Q. (2016) conducted a meta-analysis of 12 studies with 1580 HCC infected HBV, 2033 infected with HBV and 1116 healthy subjects, found that genotypes contain A allele of TNF-α -308 increased HCC risk compared to the genotype GG, when using the control group is healthy people. The AA genotype increases HCC risk when compared to the GG genotype, when using control group is infected with HBV (OR = 2,773; 95% CI = 1,107 - 6,945; p=0,03). Currently, there are a lot of subclinical and clinical evidences above relationship between TNF-α-308 polymorphism and HCC risk. In fact, the formation of HCC usually occurs after a prolonged course of chronic hepatitis so that factors that control inflammatory processes such as TNF-α greatly influence these activities. Although TNFα is a multifunctional cytokine, have inflammatory effect, against viral and bacterial, but the uncontrolled increase in TNFα levels causes angiogenesis to promote cancer formation through its effects on angiogenesis agents, oxidative radicals. That is the cause the individuals carrying A allele of TNFα -308, are more involved in TNFα production, will be more susceptible to HCC formation. 4.3. TGF-β1-509 C>T polymorphism 4.3.1. The frequency of allele and genotypes of TGF-β1- 509 C>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 <0.05 4.3.2. TGF-β1-509 C>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 8 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. Fatlle