GATA3通过调控LIFR抑制人乳腺癌细胞的迁移能力OACSTPCD
GATA3 Inhibits Migration of Human Breast Cancer Cells by Regula-ting LIFR
目的 探究GATA结合蛋白3(GATA binding protein 3,GATA3)对乳腺癌细胞迁移能力的影响.方法 在MCF7 细胞中利用慢病毒载体介导的基因干涉技术敲低 GATA3 基因,使用实时定量荧光 PCR(qRT-PCR)和蛋白质印迹检测GATA3 和LIFR的mRNA和蛋白表达水平,Transwell实验检测MCF7 细胞的迁移能力.在MCF7 和T47D细胞中用染色质免疫沉淀(ChIP-qPCR)实验检测GATA3 在LIFR的启动子区的结合位点.在敲低GATA3 基因的MCF7 细胞中回补LIFR,通过细胞划痕实验和Transwell实验检测MCF7细胞的迁移能力.结果 与对照组相比,敲低GATA3 基因的MCF7 细胞的迁移能力增强(P均<0.05).与对照组相比,敲低 GATA3 基因的 MCF7 细胞的 LIFR表达水平降低(P均<0.05).乳腺癌细胞 MCF7 与T47D中GATA3 在LIFR的启动子区有结合(P均<0.05).在敲低GATA3 基因的MCF7 细胞中稳定过表达LIFR可以部分挽救GATA3 基因敲低引起的细胞迁移能力的增强(P均<0.05).结论 GATA3 通过转录激活LIFR抑制乳腺癌细胞MCF7 的迁移.
Objective To explore the effect of GATA binding protein 3 on the migration ability of breast cancer cells.Methods Lentivirus-mediated GATA3-knockdown cell line was established to study the expression levels and function of GATA3 by performing real-time quantitative fluorescent PCR,Western blotting and Transwell assay in vitro.The binding sites of GATA3 in LIFR promoter region was detected by Chromatin immunoprecipitation assay(ChIP-qPCR)assay in MCF7 and T47D cells.LIFR was overexpressed in MCF7 cells with reduced GATA3 expression,and the migra-tion capacity of MCF7 cells was measured by Wound healing assay and Transwell assay.Results Compared with the control group,the group of MCF7 cells that knocked down GATA3 had enhanced migration ability(all P<0.05),and decreased expression of LIFR(all P<0.05).ChIP-qPCR data showed a physical binding of GATA3 on the promoter region of LIFR in MCF7 and T47D cells(all P<0.05).Overexpression of LIFR rescued the enhanced cell migration induced by depletion of GATA3 in MCF7 cells(all P<0.05).Conclusion GATA3 inhibits the migration of breast cancer cells MCF7 through transcriptional activation of LIFR.
张璐;张瑞;刘俊;杨安钢
新乡医学院医学技术学院 河南省新乡市,453003空军军医大学基础医学院免疫学教研室 西安市,710032新乡医学院医学技术学院 河南省新乡市,453003||空军军医大学基础医学院免疫学教研室 西安市,710032
临床医学
乳腺癌GATA结合蛋白3白血病抑制因子受体LIFR细胞迁移
breast cancerGATA binding protein 3leukemia inhibitory factor receptorcell migration
《医学分子生物学杂志》 2024 (004)
293-299 / 7
国家重点实验室自主研究课题(No.CBSKL2022ZZ04) This work was supported by a grant from the Independent Research Project of State Key Laboratory(No.CBSKL2022ZZ04)
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