Li Jiaying, Wu Hongjiao, Gao Hui, Kou Ruihuan, Xie Yuning, Zhang Zhi, Zhang Xuemei
School of Public Health, North China University of Science and Technology, Tangshan, China.
College of Life Science, North China University of Science and Technology, Tangshan, China.
PeerJ. 2021 Feb 1;9:e10754. doi: 10.7717/peerj.10754. eCollection 2021.
Toll-like receptor 4 (TLR4), as a key regulator of both innate and acquired immunity, has been linked with the development of various cancers, including esophageal cancer. This study aims to analyze the association of potential functional genetic polymorphisms in TLR4 with the risk of esophageal cancer.
This case-control study involved in 480 ESCC patients and 480 health controls. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype TLR4 rs1927914 polymorphism. Taqman probe method was used to determine the genotypes of TLR4 rs11536891 and rs7873784 variants. The relationship between TLR4 genetic variation and ESCC risk was analyzed by Logistic regression model by calculating the odds ratio () and 95% confidence interval (95% CI).
Compared with TLR4 rs1927914 AA genotype carriers, GG carriers had a lower ESCC risk (OR = 0.59, 95% CI [0.38-0.93], = 0.023). Stratification analysis by age showed that TLR4 rs1927914 GG could affect the risk of ESCC in elderly people (OR = 0.59, 95% CI [0.36-0.97]). Smoking stratification analysis indicated that rs1927914 GG carriers were related to ESCC susceptibility among non-smokers ( = 0.36, 95% CI [0.18-0.73]). Dual luciferase reporter assay suggested that rs1927914 G-containing TLR4 promoter displayed a 1.76-fold higher luciferase activity than rs1927914 A-containing counterpart in KYSE30 cells. Electrophoretic mobility shift assay (EMSA) showed the KYSE30 cell nuclear extract was able to bind the probe with rs1927914 G allele and this DNA-protein interaction could be eliminated by competition assays with unlabeled rs1927914 G probe, which indicating that the binding is sequence-specific. Our results also showed that TLR4 rs7873784 (G>C) and rs11536891 (T>C) conformed to complete genetic linkage. The genotype distributions of TLR4 rs11536891 variant among ESCC patients and normal controls have no statistical significance.
The TLR4 rs1927914 variant contributes to the ESCC risk by effecting the promoter activity.
Toll样受体4（TLR4）作为先天性和获得性免疫的关键调节因子，与包括食管癌在内的多种癌症的发生发展有关。本研究旨在分析TLR4潜在功能性基因多态性与食管癌风险的关联。
本病例对照研究纳入480例食管癌患者和480例健康对照。采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法对TLR4 rs1927914多态性进行基因分型。采用Taqman探针法确定TLR4 rs11536891和rs7873784变异的基因型。通过计算比值比（OR）和95%置信区间（95%CI），采用Logistic回归模型分析TLR4基因变异与食管癌风险的关系。
与TLR4 rs1927914 AA基因型携带者相比，GG携带者的食管癌风险较低（OR = 0.59，95%CI [0.38 - 0.93]，P = 0.023）。按年龄分层分析显示，TLR4 rs1927914 GG可影响老年人的食管癌风险（OR = 0.59，95%CI [0.36 - 0.97]）。吸烟分层分析表明，rs1927914 GG携带者与非吸烟者的食管癌易感性有关（OR = 0.36，95%CI [0.18 - 0.73]）。双荧光素酶报告基因检测表明，在KYSE30细胞中，含rs1927914 G的TLR4启动子的荧光素酶活性比含rs1927914 A的对应物高1.76倍。电泳迁移率变动分析（EMSA）显示，KYSE30细胞核提取物能够与含rs1927914 G等位基因的探针结合，并且这种DNA-蛋白质相互作用可以通过与未标记的rs1927914 G探针的竞争试验消除，这表明这种结合是序列特异性的。我们的结果还表明，TLR4 rs7873784（G>C）和rs11536891（T>C）符合完全遗传连锁。TLR4 rs11536891变异在食管癌患者和正常对照中的基因型分布无统计学意义。
TLR4 rs1927914变异通过影响启动子活性影响食管癌风险。
