Lentiviral vector for E6AP small-interfering RNA and its effect on breast cancer growth

VernacularTitle:慢病毒载体介导E6AP基因敲低乳腺癌细胞株的构建及其功能检测
Author: Zhida CHEN ; Lihua DING ; Jie LIU ; Yanan ZHANG ; Jie ZHU ; Xiaoli LUO ; Guoxiao LIU ; Qinong YE ; Bo WEI
Publication Type:Journal Article
Keywords: E6AP; RNA,small interfering; gene expression; breast neoplasms
From: Military Medical Sciences 2014;(11):863-866,870
CountryChina
Language:Chinese
Abstract: Objective To construct the lentiviral vector (pSIH-H1) for E6AP small-interfering RNA(siRNA) and to detect its effect on breast cancer ZR 75-1cell growth.Methods E6AP siRNA was designed and constructed based on hu-man papillomavirus E6-associated protein ( E6AP) cDNA sequence.The expression of E6AP was examined by real-time quantitative PCR(qRT-PCR) and Western blotting.The effect of E6AP on ZR75-1 cell growth was determined by cck-8 kit.Results DNA sequencing indicated that E 6AP siRNA expression vector was constructed successfully .qRT-PCR and Western blotting experiments showed that pSIH-H1-E6AP siRNA could suppress the E6AP gene expression.Suppression of E6AP could markedly inhibit the growth of ZR 75-1.Conclusion A lentivirus RNA interference ( RNAi) vector targeting E6AP gene is successfully constructed ,which inhibits the cell growth of ZR 75-1.