miR-21靶向PDCD4调控HPV16+及HPV18+宫颈癌细胞增殖的实验研究  

Study on MiR-21 Regulating the Proliferation of HPV16+and HPV18+Cervical Cancer Cells via Targeting PDCD4

作　　者：刘文博[1] 陈玲玲[1] 曹丽娟[1] LIU Wenbo;CHEN Lingling;CAO Lijuan(Chengde Central Hospital,Chengde 067000,China)

机构地区：[1]承德市中心医院,承德067000

出　　处：《病毒学报》2020年第6期1109-1114,共6页Chinese Journal of Virology

摘　　要：高危型人乳头瘤病毒(Human papillomavirus,HPV)感染是宫颈癌发生的危险因素,微小RNA(microRNA,miR)-21在宫颈癌中表达增多且与高危型HPV载量呈正相关,抑制miR-21能够靶向程序性死亡因子4(Programmed cell death 4,PDCD4)抑制肝癌细胞的增殖,但抑制miR-21对高危型HPV感染宫颈癌细胞增殖的调控作用及机制未阐明。因此,为研究miR-21靶向PDCD4调控HPV16+及HPV18+宫颈癌细胞增殖的作用,本研究培养了HPV16+Siha细胞及HPV18+HeLa细胞,转染阴性对照(Negative control,NC)抑制物、miR-21的抑制物、NC siRNA、PCDC4 siRNA后检测细胞活力及miR-21、PDCD4、β-catenin、cyclinD1的表达,验证miR-21对PDCD4的靶向调控。结果显示,转染miR-21抑制物后,Siha细胞和HeLa细胞的增殖活力及细胞中β-catenin、cyclinD1的表达均降低,细胞中PDCD4的表达增加;转染miR-21模拟物后,含有PDCD4基因3′UTR的双荧光素酶报告基因的荧光活力降低;转染PDCD4 siRNA后,miR-21抑制物抑制细胞活力、增加细胞PDCD4表达的作用被逆转。本研究提示,miR-21能够在HPV16+及HPV18+宫颈癌细胞靶向抑癌基因PDCD4,抑制miR-21表达能够增加PDCD4表达并抑制细胞增殖。Human papillomavirus(HPV)infection is a major risk factor for cervical cancer.Expression of microRNA(miR)-21 in cervical cancer is increased and correlated positively with high HPV load.Inhibition of miR-21 could inhibit the proliferation of hepatoma cells by targeting programmed cell death 4(PDCD4).However,the regulatory effect and mechanism of inhibition of miR-21 on proliferation of HPV-infected cervical cancer cells are not known.We studied how miR-21 regulates the proliferation of HPV16+and HPV18+cervical cancer cells by targeting PDCD4.We cultured HPV16+Siha cells and HPV18+HeLa cells.Then,we transfected them with negative control(NC)inhibitors,miR-21 inhibitors,NC small interfering(si)RNA and PDCD4 siRNA.Next,we tested cell viability as well as expression of miR-21,PDCD4,β-catenin,cyclinD1,and verified the effect of miR-21 on PDCD4 regulation.The proliferative ability of SiHa cells and HeLa cells,expression ofβ-catenin and cyclinD1 in Siha cells and HeLa cells decreased,and PDCD4 expression increased after transfection with a miR-21 inhibitor.Fluorescence of a double-luciferase reporter gene containing the 3′UTR of PDCD4 decreased after transfection with a miR-21 mimic.The effects of the miR-21 inhibitor on decreasing cell viability and increasing PDCD4 expression were reversed.Our data suggest that miR-21 can target the tumor-suppressor gene PDCD4 in HPV16+and HPV18+cervical cancer cells,and that inhibiting miR-21 expression can increase PDCD4 expression and inhibit proliferation of cancer cells.

关 键 词：宫颈癌 微小RNA(miR)-21 高危型HPV 程序性死亡因子4(PDCD4) 增殖 

分 类 号：R373.9[医药卫生—病原生物学]