HPV感染宫颈组织中14-3-3σ表达及启动子甲基化水平检测

目的:检测HPV感染宫颈组织中14-3-3σ启动子甲基化与14-3-3σmRNA的表达。方法:采用甲基化特异性PCR技术检测26例宫颈CIN3+宫颈原位鳞癌,39例宫颈尖锐湿疣及35例对照标本中14-3-3σ基因启动子区CpG岛甲基化状态,Q-PCR法检测14-3-3σmRNA表达,Western Blot检测14-3-3σ蛋白的表达。结果:宫颈CIN3+宫颈原位鳞癌组中14-3-3σ基因启动子区CpG岛甲基化率为73.08%,明显高于对照组的22.86%,差异有统计学意义(P0.05);尖锐湿疣组为17.94%与对照组比较,差异无统计学意义(P0.05)。宫颈CIN3+宫颈原位鳞癌组的14-3-3σmRNA及蛋白表达较对照组减少,宫颈尖锐湿疣组14-3-3σmRNA及蛋白表达较对照组增加。结论:HPV感染宫颈CIN3+宫颈原位鳞癌组织中14-3-3σ表达减少可能与14-3-3σ启动子高甲基化相关。

Expression of 14-3-3σand the detection of methylation of 14-3-3σgene promoter in the cervix with HPV infection

Objective:To detect the level of methylation of 14-3-3σ gene promoter region, and the ex-pression of 14-3-3σ mRNA and protein in cervix with HPV infection. Methods:The levels of methylation of 14-3-3σ gene promoter CPG island,and the expression of 14-3-3σ mRNA and protein were detected by methylation-specific PCR, Q-PCR and western-blot respectively in 39 cases with cervical condyloma accumi-natum ( CA) , 35 controls and 26 cases with cervical intraepithelial neoplasia ( CIN3) complicated with cervi-cal squamous cell carcinoma ( SCC) . Results:The rate of the methylation of 14-3-3σgene promoter CPG is-land in the samples of SCC was 73.08%, which was higher than that in control group (22.86%), with a sig-nificant difference (P<0.05). There was no significant difference in the rate between the samples of CA (17.94%) and of controls (P>0.05). The expression of 14-3-3σ mRNA and protein in the patients with CIN3 complicated with SCC was lower than that in controls (P<0.05). The expression of 14-3-3σ mRNA and protein in the patients with CA was higher than those in controls (P<0.05). Conclusion: The reduced expression of 14-3-3σin the patients with CIN3 complicated with SCC may be associated with the hypermeth-ylation of 14-3-3σ gene promoter region.