CAJ | 학술논문

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。
목적：채용 shRNA 침묵 Piwil2기인적표체，탐토기대궁경암세포주증식화쇠로특성적영향。방법통과지질체 Lipofectamine2000전염기술화표령매소사선，건립은전Sh-piwil2궁경암세포주，채용실시형광정량역전록취합매련반응(qRT-PCR)화 Western blot 법험증침묵효과。채용세포증식실험、집락형성실험、세포주기화세포쇠로등실험관찰침묵 Piwil2표체대궁경암세포생물학특성적영향。결과건립은정전염 Sh-piwil2궁경암세포주，여대조조상비，침묵Piwil2표체후세포적증식화집락형성능력균현저강저(P <0．05)；세포 G0/ G1기비례증가、S 기비례명현하강( P <0．05)，동시침묵 Piwil2후세포쇠로수량명현증다( P <0．05)。결론침묵 Piwil2기인가발휘항궁경암작용，추측Piwil2기인가작위궁경암림상치료적일개잠재파점。
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.