摘要目的 探讨E钙黏蛋白(CDH1)基因3'-非翻译区终止密码子下游54 bp处C/T单核苷酸多态性(3'-UTR+54C/T SNP)与宫颈癌的易感性.方法 构建含CDH1基因3'-UTR+54C/TSNP DNA序列的荧光素酶表达载体,利用双荧光素酶报告基因检测系统观察CDH1基因3'-UTR+54C/T SNP转染后人胚肾细胞株293T细胞的荧光素酶活性(RLA);采用PCR-限制性片段长度多态性(RFLP)方法检测280例宫颈癌患者(病例组)和330例健康妇女(对照组)的CDH1基因3'-UTR+54C/T SNP的基因型及等位基因频率分布,进一步分析其与宫颈癌易感性的关系.结果 双荧光素酶报告基因检测系统观察显示,转染CDH1基因3'-非翻译区(3'-UTR)C等位基因后293T细胞的RLA平均为1.46,转染CDHl基因3'-UTR T等位基因后293T细胞的RLA平均为3.01,两者比较,差异有统计学意义(t=2.94,P=0.042).PCR-RFLP方法检测显示,CDH1基因3'-UTR C等位基因频率病例组为80.7%,明显高于对照组的74.5%(χ2=6.59,P=0.010).病例组T/T、T/C、C/C基因型频率分别为4.3%、30.0%、65.7%,对照组分别为5.8%、39.4%、54.8%,两组间比较.差异有统计学意义(χ2=7.45,P=0.024).与T/T或T/C基因型比较,携带C/C基因型者宫颈癌的发病风险明显增加(OR=1.578,95%CI=1.136～2.191).结论 CDH1基因3'-UTR C等位基因可能降低荧光素酶报告基因的表达;C/C基因型可能是宫颈癌发病的潜在危险因素.

abstractsObjective To investigate the effect of CDH1 3'-UTR + 54C/T single nucleotide polymorphism(SNP) on expression of luciferase reporter gene and its association with susceptibility to cervical cancer. Methods The luciferase gene expression vectors containing CDH1 3'-UTR +54C/T SNP C or T allelotype were constructed. The effect of CDH1 3'-UTR + 54C/T SNP on expression of luciferase reporter gene in 293 T cells were tested by daul lucfferase reporter assay system. The CDH1 3'-UTR + 54C/ T SNP was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 280 cervical cancer patients and 330 healthy controls. Results The lucfferase activity analysis showed that the relative luciferase activity (RLA) of 293T cells with C allelotype was 1.46, which was significantly lower than that of the 293 T cells with T allelotype (3.01; t=2. 94, P =0. 042). There was no significant difference in age distribution between the cervical cancer patients and the healthy controls. The genotype frequency distribution of CDH1 3 '-UTR + 54C/T SNP in healthy controls did not significantly differ from that expected by Hardy-Weinberg equilibrium (P>0.05). The C allelotype frequency of CDH1 in cervical cancer patients was 80. 7%, which was significantly higher than that in healthy controls (74. 5%;χ2 =6.59, P=0.010). The T/T, T/C and C/C genotype frequencies of cervical cancer patients and healthy controls were 4. 3%, 30. 0%, 65. 7% and 5. 8%, 39. 4%, 54. 8%, respectively, which were significandy different (χ2=7.45, P =0.024). Compared with individuals with T/T or T/C genotypa, individuals with C/C genotype had significantly higher risks of developing cervical cancer (OR = 1. 578,95%CI=1.136 -2.191). Conclusion The C allelotypa of CDH1 3'-UTR + 54C/T SNP might decrease the expression of lucfferase reporter gene and the C/C genotypa might be a potential risk for cervical cancer development.