早期生长反应基因在宫颈癌中的表达及与高危型人乳头瘤病毒感染的关系

Expression of early growth response gene in cervical cancer and the relationship with high-risk human papillomavirus infection

目的探讨早期生长反应基因(Egr-1)在宫颈癌中的表达及与高危型人乳头瘤病毒(HPV)感染的关系。方法选取2018年6月-2019年6月间在宁波市鄞州人民医院行手术治疗的宫颈癌及癌前病变患者192例,另收集同期行手术治疗的宫颈良性病变患者52例。免疫组化法检测宫颈组织中Egr-1表达情况。培养人宫颈癌细胞株C-33A(HPV-)、CaSki(HPV16+)、HeLa299(HPV18+),RT-PCR和Western blot检测Egr-1 mRNA和蛋白表达情况,并使用HPV16 E6、HPV18 E6特异性siRNA序列沉默CaSki、HeLa299细胞中E6表达,检测对Egr-1 mRNA和蛋白表达的影响。结果Egr-1阳性表达率在宫颈癌组、宫颈上皮内瘤变组、宫颈良性病变组中差异有统计学意义(P<0.01);其中宫颈癌组Egr-1表达阳性率明显低于宫颈上皮内瘤变组,宫颈上皮内瘤变组Egr-1表达阳性率明显低于宫颈良性病变组。宫颈癌细胞株HeLa299、CaSki中Egr-1 mRNA和蛋白表达水平均明显高于宫颈癌细胞株C-33A(P<0.01);Egr-1 mRNA和蛋白表达水平在宫颈癌细胞株CaSki、HeLa299之间差异无统计学意义(P>0.05)。转染48 h后,siRNA组CaSki、HeLa299细胞中E6 mRNA和蛋白表达水平均明显低于空白对照组、阴性对照组(P<0.05);siRNA组CaSki、HeLa299细胞中Egr-1 mRNA和蛋白表达水平明显高于空白对照组、阴性对照组(P<0.05)。结论宫颈癌组织中Egr-1低表达;高危型HPV16、18可能通过E6蛋白参与的信号通路下调Egr-1表达,提升宫颈癌细胞活性。

Objective To explore the expression of early growth response(Egr-1)gene in cervical cancer and the relationship with high-risk human papillomavirus(HPV)infection.Methods From June 2018 to June 2019,192 patients with cervical cancer and cervical precancerous lesion treated by surgery in Yinzhou People's Hospital were selected,and 52 patients with cervical benign lesions treated by surgery were collected.Immunohistochemical method was used to detect the expression of Egr-1 in cervical tissue.Human cervical cancer C-33A(HPV-),CaSki(HPV16+),and HeLa299(HPV18+)cells were cultured.RT-PCR and Western blot were used to detect expressions of Egr-1 mRNA and protein.HPV16 E6,HPV18 E6 specific siRNA sequences were used to silence E6 expressions in CaSki and HeLa299 cells.The effect on expressions of Egr-1 mRNA and protein was detected.Results There was statistically significant difference in the positive expression rate of Egr-1 among cervical cancer group,cervical intraepithelial neoplasia(CIN)group,and cervical benign lesion group(P<0.01),the positive expression rate of Egr-1 in cervical cancer group was significantly lower than that in CIN group,the positive expression rate of Egr-1 in CIN group was significantly lower than that in cervical benign lesion group.The expression levels of Egr-1 mRNA and protein in cervical cancer HeLa299 and CaSki cells were statistically significantly higher than those in cervical cancer C-33A cells(P<0.01).There was no statistically significant difference in the expression levels of Egr-1 mRNA and protein between cervical cancer CaSki cells and cervical cancer HeLa299 cells(P>0.05).After 48 hours of transfection,the expression levels of E6 mRNA and protein in cervical cancer CaSki and HeLa299 cells in siRNA group were statistically significantly lower than those in blank control group and negative control group(P<0.05);the expression levels of Egr-1 mRNA and protein in cervical cancer CaSki and HeLa299 cells in siRNA group were statistically significantly lower than those in blank control group and negative control group(P<0.05).Conclusion Egr-1 expression level is low in cervical cancer tissue,HPV16 and HPV18 may downregulate Egr-1 expression and improve cervical cancer cell activity via E6 protein.