GATA3通过调控LIFR抑制人乳腺癌细胞的迁移能力

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 migration 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.