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目的 Toll样受体7(toll like receptor 7,TLR7)在识别和介导抗丙型肝炎病毒(hepatitis C virus,HCV)免疫应答中有重要作用。本研究旨在分析TLR7 rs179016单核苷酸多态性对TLR7基因转录活性的影响。方法收集22例健康男性者血液,提取全基因组DNA及外周血单个核细胞,采用实时定量逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)法检测TLR7 mRNA水平,并采用荧光素酶报告基因法,检测含有rs179016 C、G等位基因的质粒及空白质粒的HEK-293T细胞在培养24 h后荧光素强度,分析该位点对TLR7转录活性的影响。结果携带rs179016危险等位基因C组TLR7 mRNA表达水平低于rs179016G组(t=-7.352,P<0.001),转染含有rs179016G质粒的细胞表达较高强度的荧光素酶活性。结论 TLR7 rs179016多态性可能通过影响TLR7基因的转录和表达水平来影响该受体功能的发挥,可能与HCV的易感性和感染转归有关。
Objective Toll-like receptor 7 (TLR7) plays an important role in recognizing and mediating the immune response against hepatitis C virus (HCV). The purpose of this study was to analyze the effect of TLR7 rs179016 SNP on TLR7 gene transcription activity. Methods Twenty-two healthy male subjects were collected to collect whole genome DNA and peripheral blood mononuclear cells. The level of TLR7 mRNA was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR) Luciferase reporter gene method, HEK-293T cells containing rs179016 C and G alleles and blank plasmids were cultured for 24 h, and the fluorescence intensity of TLR7 was detected. Results Compared with rs179016G group (t = -7.352, P <0.001), the expression of TLR7 mRNA in the rs179016 risk allele group was lower than that in the rs179016G group. The cells transfected with the rs179016G plasmid expressed higher luciferase activity. Conclusion The polymorphism of TLR7 rs179016 may affect the function of TLR7 gene by affecting the transcription and expression of TLR7 gene, which may be related to the susceptibility of HCV and the outcome of infection.