沉默 Piwil2表达对子宫颈癌细胞增殖和衰老的影响

Silencing of Piwil2 expression on the proliferation and senescence of cervical cancer cells

目的采用shRNA沉默Piwil2基因的表达,探讨其对宫颈癌细胞株增殖和衰老特性的影响。方法通过脂质体Lipofectamine2000转染技术和嘌呤霉素筛选,建立稳转Sh-piwil2宫颈癌细胞株,采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)和Western blot法验证沉默效果。采用细胞增殖实验、集落形成实验、细胞周期和细胞衰老等实验观察沉默Piwil2表达对宫颈癌细胞生物学特性的影响。结果建立稳定转染Sh-piwil2宫颈癌细胞株,与对照组相比,沉默Piwil2表达后细胞的增殖和集落形成能力均显著降低(P<0.05);细胞G0/G1期比例增加、S期比例明显下降(P<0.05),同时沉默Piwil2后细胞衰老数量明显增多(P<0.05)。结论沉默Piwil2基因可发挥抗宫颈癌作用,推测Piwil2基因可作为宫颈癌临床治疗的一个潜在靶点。

Objective To explore the effect of silencing Piwil2 gene by shRNA on the proliferation and senescence of cervical cancer cells. Methods Constructed the stable Sh-piwil2 cervical cancer cells via Lipofectamine2000 mediation and puromycin selection. The efficiency of transfection was confirmed by real-time fluorescence quantita-tive polymerase chain reaction（qRT-PCR） and Western blot. Cell proliferation assay, colony forming assay, cell cycle assay and cell senescence assay were detected the influence of silencing Piwil2 gene on the biological behavior of cervical cancer cells. Results The stable Sh-piwil2 cervical cancer cells were constructed. Compared with the Vector groups, the growth and colony forming rate were significantly decreased. Moreover, the cell cycle was arres-ted in the G0 / G1 phase and the cell senescence was increased through silence the expression of Piwil2 gene. Con-clusion Silencing Piwil2 gene can effectively suppress cell proliferation of cervical cancer cells. We speculate that Piwil2 gene may become a potential target in clinical therapy for cervical cancer.