人乳头瘤病毒16 E6基因对子宫颈癌SiHa细胞株中E-钙黏蛋白表达的影响

Effect of human papillomavirus 16 E6 gene on the expression of E-cadherin in human cervical cancer cell line SiHa

目的 探讨人乳头瘤病毒16 E6（HPV16 E6）基因对子宫颈癌SiHa细胞株中E-钙黏蛋白（E-cad）表达的影响.方法 在子宫颈癌SiHa细胞中运用瞬时转染体系建立HPV16 E6过表达体系,采用四甲基偶氮唑盐（MTT）法检测SiHa细胞增殖情况,采用反转录聚合酶链反应（RT-PCR）法及蛋白印迹法分别检测HPV16 E6转染后细胞中E-cad mRNA及蛋白表达情况.结果 与空载体（加入pcDNA3.1）组和空白对照（未转染质粒）组比较,E6组（pcDNA3-1-HPV16 E6）的细胞存活率更高（均P〈0.05）,空白对照组和空载体组比较,差异无统计学意义（P〉0.05）.E6组、空载体组与空白对照组的E-cad mRNA相对表达量分别为0.26±0.12、0.82±0.14、0.83±0.21,蛋白相对表达量分别为0.62±0.02、1.33±0.04、1.31±0.05.E6组的E-cad mRNA与蛋白表达水平低于另外两组,差异均有统计学意义（均P〈0.05）,但空白对照组和空载体组相比,差异无统计学意义（均P〉0.05）.结论 HPV16 E6基因瞬时转染过表达能降低子宫颈癌细胞中E-cad的表达,与子宫颈癌的发生、发展密切相关.

Objective To investigate the effect of human papillomavirus （HPV） 16 E6 gene on the expression of E-cadherin （E-cad） in cervical cancer cell line SiHa. Methods HPV16 E6 expression system was established in the cervical cancer cell line SiHa by using transient transfection system, MTT method was used to detect SiHa cell proliferation activity, and reverse transcription-polymerase chain reaction （RT-PCR） and western blot method was used respectively to detect E-cad mRNA and protein expression level in cells after HPV16 E6 transfection. Results Compared with the blank control group （non-transfected plasmid） and the vector group （the addition of pcDNA3.1）, the cell viability rate of the E6 group （pcDNA3-1-HPV16 E6） was significantly increased （P〈0 .05）, while there was no significant difference between the vector group and the blank control group （P〉0.05）. The relative expressions of E-cad mRNA in the E6 group, the vector group and the blank control group were 0.26±0.12, 0.82±0.14, 0.83±0.21 respectively, then the protein relative expressions in the three groups were 0.62±0.02, 1.33±0.04, 1.31±0.05 respectively. The expressions level of E-cad mRNA and protein in E6 group were significantly lower than those in the other two groups （all P〈0.05）. However, there was no significant difference between the vector group and the blank control group （both P〉0.05）. Conclusion The instantaneous transfection of HPV16 E6 gene can reduce the expression of E-cad in cervical cancer cells, which is closely related to the occurrence and development of cervical cancer.