摘要
目的通过体外实验探讨沉默PIK3R2基因对宫颈癌细胞增殖、迁移及侵袭的影响和机制。方法qRT-PCR和Western blot技术检测正常宫颈细胞Ect1/E6E7和4种宫颈癌细胞(C33A、HeLa、SiHa和CaSki)中PIK3R2的表达。将携带PIK3R2 shRNA的慢病毒载体转染SiHa细胞,建立稳定沉默PIK3R2的细胞株LV-PIK3R2-shRN,MTT法检测细胞增活力,Transwell实验检测细胞的迁移和侵袭能力,Western blot检测增殖相关基因(CyclinD1和p21)、迁移侵袭相关基因(MMP-2和MMP-9)及PI3K/Akt信号通路重要蛋白(PI3K、p-PI3K、Akt和p-Akt)的表达水平。结果与正常宫颈细胞Ect1/E6E7相比,4种宫颈癌细胞C33A、HeLa、SiHa、CaSki中PIK3R2 mRNA和PIK3R2蛋白相对表达量显著升高,差异具有统计学意义(P<0.05)。与LV-NC组相比,LV-PIK3R2-shRNA组SiHa细胞PIK3R2蛋白的表达显著降低,SiHa细胞在24h、48h、72h和96h时间点细胞活力显著降低,差异具有统计学意义(P<0.05)。与LV-NC组相比,LV-PIK3R2-shRNA组SiHa细胞的迁移和侵袭细胞数目显著降低,差异具有统计学意义(P<0.05)。与LV-NC组比较,LV-PIK3R2-shRNA组p-PI3K和p-AKT蛋白的表达显著降低,差异具有统计学意义(P<0.05)。与LV-NC组相比,LV-PIK3R2-shRNA组SiHa细胞的增殖活力显著降低,迁移和侵袭细胞数目显著减少差异具有统计学意义(P<0.05);与LV-PIK3R2-shRNA组相比,LV-PIK3R2-shRNA+激动剂组SiHa细胞的增殖活力显著升高,迁移和侵袭细胞数目显著增多,差异具有统计学意义(P<0.05)。结论沉默PIK3R2可抑制PI3K/AKT信号通路激活,进而抑制宫颈癌细胞的增殖、迁移和侵袭。
Objective To investigate the effect of PIK3R2 gene silencing on the proliferation,migration and invasion of cervical cancer cells in vitro and it mechanism.Methods The expression of PIK3R2 in normal cervical cells Ect1/E6E7 and four cervical cancer cells(C33A,HeLa,SiHa and CaSki)wasdetected by qRT-PCR and Western blot.The lentiviral vector carrying PIK3R2 shRNA was transfected into SiHa cells,and the cell line LV-PIK3R2-shRN stably silencing PIK3R2 was established.The cell viability was detected by MTT assay.The migration and invasion ability of cells were detected by Transwell assay.The expression levels of proliferation-related genes(CyclinD1 and p21),migration and invasion related genes(mmp-2 and mmp-9)and important proteins of PI3K/Akt signaling pathway(PI3K,p-pI3K,Akt and p-AKT)were detected by Western blot.Results The ex-pression of PIK3R2 was significantly up-regulated in four cervical cancer cells compared to Ect1/E6E7(P<0.05).The cell line stably silencing PIK3R2 was successfully constructed,and the expression level of PIK3R2 protein was significantly decreased(P<0.05).Silencing PIK3R2 inhibited the proliferation,migration and invasion of SiHa cells and inhibited the activation of PI3K/AKT signaling pathway(P<0.05).The activation of PI3K/AKT signa-ling pathway reverses the inhibitory effect of PIK3R2 silencing on the proliferation,migration and invasion of SiHa cells(P<0.05).Conclusions Silencing PIK3R2 inhibits the activation of PI3K/AKT signaling pathway,thereby inhibiting the proliferation,migration and invasion of cervical cancer cells.
作者
周业娣
成晓燕
钱琳玉
ZHOU Yedi;CHENG Xiaoyan;QIAN Linyu(Department of Gynecology and Obstetrics,Nantong Maternal and Child Health Hospital,Nantong 226000,Jiangsu,China)
出处
《中国性科学》
2020年第9期16-20,共5页
Chinese Journal of Human Sexuality
关键词
慢病毒
PIK3R2
宫颈癌
细胞增殖
迁移
侵袭
Lentivirus
PIK3R2
Cervical cancer
Cell proliferation
Migration
Invasion
作者简介
通讯作者:周业娣,E-mail:303403622@qq.com。
