| Hepatocellular carcinoma is an important malignant tumor in China, the occurrence ofit is affacted by many factors, including viral and the genetic risk factors. In China, chronichepatitis B virus infection is the major risk factor for HCC. Recent studies have confirmedthat the occurrence of HCC is related with the chronic liver inflammation stimuli followedby HBV infection. But, the majority of patients with chronic HBV infection don’t developinto HCC, indicating that the genetic susceptibility of Chinese plays a critical role in thedevelopment of HCC.MicroRNAs (miRNAs) are very important small non-coding regulatory RNAs thatcan play critical roles throughout cell proliferation and differentiation, development andapoptosis. There is a close relationship between miRNApolymorphism and thedevelopment of HCC. Single nucleotide polymorphism (SNP) within the miRNAseedregion which can change the miRNAmature and the binding affinity with the target gene,is related with tumor susceptibility. The miR-34family is a very important non-codingRNAs in the development of cancer, including HCC. As the direct target of p53, miR-34can inhibit the cell growth, induce cell apoptosis. Taken together, it has a great significanceto study the polymorphism of miR-34and HBV mutations and the interaction betweenthem in the development of HCC.Objective:This study was aimed to explore the associations of virus and genetics (the miR-34b/cpolymorphism, rs4938723) and their interactions in chronic hepatitis B (CHB), Livercirrhosis (LC), and HCC. To further elucidate the role of these factors during thedevelopment of HBV-related HCC, and reveal the viral and genetic risk factors for HCC,and then provide important clues for the prevention of the risk factors of HCC in China.Methods:This study include1012healthy controls,316Asymptomatic HBsAg carriers (ASCs),316patients with CHB,316patients with LC, and1021HCC cases. The polymorphisms ofmiR-34(rs4938723) were genotyped by Fluorescent probe-Real time quantitative PCR.Hardy-Weinberg equilibrium test was performed on the internet (http://ihg. gsf.de/ihg/snps. html). The SPSS16.0was used to analyze the data. t test for independentsample, chisquare test, analysis of variance, SNK-q test and unconditional logistic regression model were used to illustrate the associations of HBV mutations andpolymorphisms of miR-34b/c and their interactions during the development of HCC.Results:1. Associations between polymorphisms of miR-34b/c (rs4938723) and susceptibility toliver disease.(1)Compared with non-LC patients with HBV infection (ASCs and CHB), patientswith CC genotype were significantly associated with a decreased risk of LC (AOR=0.60，95%CI=0.36-0.99). But, this value is near the threshold, it should be noted that whetherthis genotype is really a protective factor for LC.(2) After stratified by gender, we found that in female:Compared with healthy control, patients with TC genotype and C allele weresignificantly associated with an increased risk of HCC (AOR=1.38，95%=1.10-1.74；AOR=1.48，95%CI=1.08-2.05). When compared with non-HCC patients with HBVinfection (ASCs, CHB and LC), patients with CC genotype and C allele were significantlyassociated with an increased risk of HCC (AOR=2.33，95%CI=1.14-4.76；AOR=1.42，95%CI=1.05-1.92).(3) After stratified by gender, we found that in male:Compared with non-LC patients with HBV infection (ASCs and CHB), patients withTC genotype and Callele were significantly associated with a decreased risk of LC(AOR=0.84，95%CI=0.71-0.99;AOR=0.74,95%CI=0.57-0.94), but, this value is near thethreshold, it should be noted that whether this genotype is really a protective factor for LC.(4) After stratified by HBV genotype, we found that in HBV genotype B:comparedwith non-LC patients with HBV infection (ASCs and CHB), patients with CC genotypewere significantly associated with a decreased risk of LC (AOR=0.11，95%CI=0.01-0.93).However, this result needs to be validated using larger sample size.(5) After stratified by HBeAg status, we found that when HBeAg was negative,compared with non-HCC patients with HBV infection (ASCs, CHB and LC), patients withCC genotype were significantly associated with an increased risk of HCC (AOR=1.69，95%CI=1.03-2.79).2. Interactions between polymorphism of miR-34b/c (rs4938273) and HBV genotypes andmutations. (1) The interaction of polymorphism of miR-34b/c and A1762T/G1764Amutation indevelopment of HCCAfter adjusted by age, HBV genotype and HBeAg status, we found that:In female, rs4938723TT genotype and “wild type” ofA1762T/G1764Aas control, thecombined effects ofTC, CC genotype, C allele and A1762T/G1764Asignificantlyincreased the risks of HCC (AOR=6.89，95%CI=2.44-19.47;AOR=18.46，95%CI=3.78-90.09;AOR=8.88，95%CI=4.61-17.09). However, there was no significantinteraction between polymorphism of rs4938723and A1762T/G1764Ain femaleparticipants.In male, rs4938723TT genotype and “wild type” ofA1762T/G1764Aas control, thecombined effects ofTC, CC genotype, C allele and A1762T/G1764Asignificantlyincreased the risks of HCC (AOR=2.99，95%CI=1.73-5.17;AOR=2.40，95%CI=1.17-4.93;AOR=3.20，95%CI=2.27-4.51). Asignificant interaction was found between rs4938723TCgenotype and A1762T/G1764Ain male participants (AOR=1.51，95%CI=1.03-2.21).(2) The interaction of polymorphism of miR-34b/c and T1674C/G mutation indevelopment of HCCAfter adjusted by age, HBV genotype and HBeAg status, we found that:In female, rs4938723TT genotype and “wild type” of T1674C/G as control, thecombined effects ofTC, CC genotype, C allele and T1674C/G significantly increased therisks of HCC (AOR=3.99，95%CI=1.48-10.75;AOR=9.50，95%CI=1.56-57.83;AOR=4.36，95%CI=2.07-9.19). However, there was no significant interaction betweenpolymorphism of rs4938723and T1674C/G in female participants.In male, rs4938723TT genotype and “wild type” of T1674C/G as control, thecombined effects ofTC, CC genotype, C allele and T1674C/G significantly increased therisks of HCC (AOR=2.47，95%CI=1.32-4.64;AOR=5.15，95%CI=1.63-16.28;AOR=3.02，95%CI=1.86-4.85). However, there was no significant interaction between polymorphismof rs4938723and T1674C/G in male participants.(3) The interaction of polymorphism of miR-34b/c and G1896Amutation in thedevelopment of HCCAfter adjusted by age, HBV genotype and HBeAg status, we found that:In female, rs4938723TT genotype and “wild type” of G1896Aas control, the combined effects ofTC, CC genotype, C allele and G1896Asignificantly increased therisks of HCC (AOR=4.86，95%CI=1.85-12.76;AOR=15.73，95%CI=2.75-90.03;AOR=5.56，95%CI=2.86-10.83). However, there was no significant interaction betweenpolymorphism of rs4938723and G1896Ain female participants.In male, rs4938723TT genotype and “wild type” of G1896Aas control, the combinedeffects of TC, CC genotype, C allele and G1896Asignificantly increased the risks of HCC(AOR=2.02，95%CI=1.15-3.57;AOR=2.86，95%CI=1.21-6.73;AOR=2.40，95%CI=1.68-3.43). However, there was no significant interaction between polymorphismof rs4938723and G1896Ain male participants.(4) The interaction of polymorphism of miR-34b/c and HBV genotype indevelopment of HCCAfter adjusted by age and HBeAg status, we found that:In female, rs4938723TT genotype and HBV genotype B as control, the combinedeffects of TC, CC genotype, C allele and HBV genotype C significantly increased the risksof HCC (AOR=2.69，95%CI=1.10-6.55;AOR=6.88，95%CI=1.54-30.75;AOR=3.30，95%CI=1.78-6.11). However, there was no significant interaction between polymorphismof rs4938723and HBV genotype C in female participants.In male, rs4938723TT genotype and HBV genotype B as control, the combinedeffects of TC, CC genotype, C allele and HBV genotype C significantly increased the risksof HCC (AOR=2.19，95%CI=1.34-3.56;AOR=2.33，95%CI=1.70-3.21;AOR=1.93，95%CI=1.05-3.53). However, there was no significant interaction between polymorphismof rs4938723and HBV genotype C in male participants.(5) The interaction of polymorphism of miR-34b/c and HBV genotype in developmentof LCAfter adjusted by age and HBeAg status, we found that:In female, rs4938723TT genotype and HBV genotype B as control, the combinedeffects of TC, CC genotype, C allele and G1896Asignificantly increased the risks of LC(AOR=2.03，95%CI=0.65-6.30;AOR=4.58，95%CI=0.54-38.90;AOR=1.91，95%CI=0.87-4.23). However, there was no significant interaction between polymorphismof rs4938723and HBV genotype C in female participants.In male, rs4938723TT genotype and HBV genotype B as control, there was no significant combined effects ofTC, CC genotype and HBV genotype C in LC (AOR=0.97，95%CI=0.48-1.95;AOR=1.47，95%CI=0.56-3.84; AOR=1.46，95%CI=0.92-2.34).However, there was a significant interaction between rs4938723TC genotype and HBVgenotype C in male participants (AOR=3.54，95%CI=1.56-8.05).3. Multivariate regression analysis for factors independently associated with the risk ofHCC and LC.(1) Independent factors contributing to HCC were evaluated in the forward stepwisemultivariate regression analysis. In addition to age, male and HBeAg negative,A1762T\G1764A(AOR=2.93，95%CI=2.07-4.16), G1896A(AOR=1.88，95%CI=1.33-2.65), T1674C/G (AOR=2.04，95%CI=1.36-3.04) were independentlyassociated with the risk of HCC.(2) Independent factors contributing to LC were evaluated in the forward stepwisemultivariate regression analysis. In addition to age,A1762T\G1764A(AOR=2.42，95%CI=1.47-3.98) and G1896A(AOR=1.69，95%CI=1.69-4.69) were independentlyassociated with the risk of LC.Conclusions:1. Age, male, HBeAg negative, T1674C/G, A1762T/G1764Aand G1896Aindependently associated with the risks of HCC.2. Age, A1762T/G1764Aand G1896Aare risk factors independently associated withthe risks of LC.3. Patients with rs4938723CC genotype are associated with decreased risk of LC. Infemale, patients who carry the TC genotype and C allele have a lower risk to develop intoLC after infected with HBV. In male, patients with TC genotype and C allele are associatedwith decreased risk of LC.4. In HBV genotype B, non-LC patients (ASCs and CHB) with CC genotype areassociated with decreased risk of LC. However, this result needs to be validated usinglarger sample size.5. When HBeAg status is negative, non-HCC patients (ASCs, CHB and LC) with CCgenotype are associated with increased risk of HCC.6. There is significant interaction betweenA1762T/G1764Amutation and polymorphism of miR-34b/c (rs4938723) on male HCC risk.7. There is significant interaction between HBV genotype and polymorphism ofmiR-34b/c (rs4938723) on male LC risk. |
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