3-疏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

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关键词：神经胶质瘤；3-疏基丙酮酸硫基转移酶；基因敲低技术

中图分类号：R739.41文献标志码：A 文章编号：2097-7174（2025）10-0934-06

DOI：10.3969/j.issn.2097-7174.2025.10.002

Abstract：Objective ∴ To construct the glioma stable cell lines GSC1023 and GBM0709 with knockdown of 3-mercapto - pyruvatesulfurtransferase （MPST），and to analyze the localization of MPST in glioma cels.Methods：Two pairs of MPST-shRNAinterferencesequences weredesignedandsynthesized，thenligatedintothepLKO.1-Puro lentiviralvector to construct the lentiviral vectorpLKO.1-MPST-shRNA for MPSTgene knockdown.This constructed vector was used to infect human glioma stem cells（GSC1023）and glioma cells（GBMo709）.Subsequently，puromycin was employed to screen and establish stable MPST-knockdown gliomacellines，namely GSC1023and GBM0709.RT-qPCR，western blot，andimmunofluorescence wereused todetect MPSTexpresion incellines.Results： GSC1023and GBM0709 stable cell lines withthe MPST gene knockdownwere successfully constructed.Theresultsof RT-qPCR showed that the expression of MPST gene in stably knockdown cellines were decreased significantly（ P<0.05 ）；the results of western blot showed that the expression of MPST in stably knockdown cellines were decreased significantly （ P<0.05 ）. The results of immunofluorescence showed thatMPSTwas mainly localized inmitochondria.Conclusion：Thestable gliomacell lines GSC1023 and GBM0709 with MPST knockdown were succesfullyestablished，laying a foundation for further exploration of the influence of MPST-mediated endogenous HS on the occurrence and development of glioma.

KeyWords：Glioblastoma；3-mercaptopyruvate sulfurtransferase；Gene knockdown techniques

神经胶质瘤是一种常见的中枢神经系统恶性肿瘤，其中以胶质母细胞瘤（Glioblastoma，GBM）侵袭性最强、致死率最高，患者确诊GBM后的平均生存时间仅为14个月[2]。（剩余10527字）