Toll样受体4基因多态性与HBV相关原发性肝癌易感性及预后的关系

Association of TLR4 gene polymorphism with susceptibility to and prognosis of hepatitis B virus-related primary liver cancer

目的探讨分析Toll样受体4(TLR4)基因多态性与HBV相关原发性肝癌易感性之间的关系以及对患者预后的影响。方法选取2015年6月-2017年7月北京中医药大学东直门医院合作的北京佑安医院收治的HBV相关肝癌患者106例作为肝癌组,同期北京中医药大学东直门医院收治的慢性乙型肝炎患者120例作为乙型肝炎组以及健康对照受试者100例作为对照组。收集各组受试者的血液标本,检测3组受试者TLR4 D299G、T399I以及A896G多态性位点基因型,并分析不同基因型肝癌患者术后复发情况。计量资料多组间比较采用方差分析,进一步两两比较采用LSD-t检验;计数资料两组比较采用χ2检验,多组间比较采用Kruskal-Wallis H秩和检验。采用非条件logistic回归模型及相对危险度近似估计值比值比(OR)估计各研究因素与HBV相关原发性肝癌发病风险的关系;生存分析绘制Kaplan-Meier曲线,生存曲线比较采用log-rank检验。结果 3组受试者TLR4 D299G位点以及TLR4 A896G位点基因型及等位基因分布差异均无统计学意义(χ2=1. 436、1. 949、1. 851、2. 599,P值均> 0. 05);而3组受试者TLR4 T399I位点基因型和等位基因比较差异均具有统计学意义(χ2=11. 654、14. 995,P值均<0. 001),其中肝癌组T等位基因频率明显高于乙型肝炎组及对照组(P <0. 05)。非条件logistic回归模型显示,TLR4 T399I CC基因型OR[95%可信区间(95%CI)]为1. 682(1. 109~2. 243),P=0. 027、TLR4 T399I TT基因型OR(95%CI)为2. 103(1. 347~2. 896),P <0. 001;TLR4 T399I T等位基因OR(95%CI)为1. 872(1. 192~2. 374),P=0. 002。TLR4 D299G位点以及TLR4 A896G位点不同基因型肝癌患者术后无复发生存情况无明显差异(χ2=0. 022、0. 471,P值均> 0. 05)。TLR4 T399I位点CC基因型肝癌患者术后无复发生存情况明显优于CT+TT基因型(χ2=6. 781,P <0. 05)。结论 TLR4 T399I位点基因多态性与HBV相关原发性肝癌易感性密切相关,该位点T基因携带可能是肿瘤发生以及预后不良的重要因素。

Objective To investigate the association of TLR4 gene polymorphism with the susceptibility to hepatitis B virus(HBV)-related primary liver cancer and its influence on the prognosis of patients. Methods A total of 106 patients with HBV-related liver cancer who were admitted to Beijing YouAn Hospital which cooperated with Dongzhimen Hospital of Beijing University of Chinese Medicine from June 2015 to July 2017 were enrolled as liver cancer group;120 patients with chronic hepatitis B who were admitted during the same period of time in Dongzhimen Hospital of Beijing University of Chinese Medicine were enrolled as hepatitis B group,and 100 healthy control subjects were enrolled as control group.Blood samples were collected from each group to detect TLR4 D299G,T399I,and A896G polymorphisms,and postoperative recurrence was analyzed for liver cancer patients with different genotypes.An analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups;the chi-square test was used for comparison of categorical data between two groups,and the Kruskal-Wallis H test was used for comparison between multiple groups.An unconditional logistic regression model and odds ratio(OR)used to estimate relative risk were used to investigate the association of each factor with the onset risk of HBV-related primary liver cancer;the Kaplan-Meier curves were plotted for survival analysis,and the log-rank test was used for comparison of survival curves between groups. Results There were no significant differences in the genotype and allele distributions of TLR4 D299G and TLR4 A896G between the three groups(χ2=1.436,1.949,1.851,and 2.599,all P>0.05),while there were significant differences in the genotype and allele distributions of TLR4 T399I between the three groups(χ2=11.654 and 14.995,both P<0.05),and the liver cancer group had a significantly higher frequency of T allele than the hepatitis B group and the control group(P<0.05).The unconditional logistic regression model showed that CC genotype of TLR4 T399I had an OR of 1.682(95%confidence interval[CI]:1.109-2.243,P=0.027),TT genotype of TLR4 T399I had an OR of 2.103(95%CI:1.347-2.896,P<0.001),and T allele of TLR4 T399I had an OR of 1.872(95%CI:1.192-2.374,P=0.002).There was no significant difference in recurrence-free survival after surgery between the liver cancer patients with different genotypes of TLR4 D299G and TLR4 A896G(χ2=0.022 and 0.471,both P>0.05).The liver cancer patients with CC genotype of TLR4 T399I had significantly better recurrence-free survival after surgery than those with CT or TT genotype(χ2=6.781,P<0.05). Conclusion TLR4 T399I gene polymorphism is closely associated with the susceptibility to HBV-related primary liver cancer,and T gene of TLR4 T399I may be an important factor for tumorigenesis and poor prognosis.