赣南医科大学学报2025，Vol.45Issue(10)：934-939,6.DOI:10.3969/j.issn.2097-7174.2025.10.002
3-巯基丙酮酸硫基转移酶敲低稳转神经胶质瘤细胞系的构建及表达
Expression and construction of stable cell lines with MPST knockdown in glioblastoma cell
摘要
Abstract
Objective:To construct the glioma stable cell lines GSC1023 and GBM0709 with knockdown of 3-mercapto-pyruvate sulfurtransferase(MPST),and to analyze the localization of MPST in glioma cells.Methods:Two pairs of MPST-shRNA interference sequences were designed and synthesized,then ligated into the pLKO.1-Puro lentiviral vector to construct the lentiviral vector pLKO.1-MPST-shRNA for MPST gene knockdown.This constructed vector was used to infect human glioma stem cells(GSC1023)and glioma cells(GBM0709).Subsequently,puromycin was employed to screen and establish stable MPST-knockdown glioma cell lines,namely GSC1023 and GBM0709.RT-qPCR,western blot,and immunofluorescence were used to detect MPST expression in cell lines.Results:GSC1023 and GBM0709 stable cell lines with the MPST gene knockdown were successfully constructed.The results of RT-qPCR showed that the expression of MPST gene in stably knockdown cell lines were decreased significantly(P＜0.05);the results of western blot showed that the expression of MPST in stably knockdown cell lines were decreased significantly(P＜0.05).The results of immunofluorescence showed that MPST was mainly localized in mitochondria.Conclusion:The stable glioma cell lines GSC1023 and GBM0709 with MPST knockdown were successfully established,laying a foundation for further exploration of the influence of MPST-mediated endogenous H2S on the occurrence and development of glioma.关键词
神经胶质瘤/3-巯基丙酮酸硫基转移酶/基因敲低技术Key words
Glioblastoma/3-mercaptopyruvate sulfurtransferase/Gene knockdown techniques分类
临床医学引用本文复制引用
王琪,李日清,陈晓霞..3-巯基丙酮酸硫基转移酶敲低稳转神经胶质瘤细胞系的构建及表达[J].赣南医科大学学报,2025,45(10):934-939,6.基金项目
广东省医学科研基金项目(B2022068) （B2022068）
