GSTP1通过调控STAT3信号通路促进乳腺癌细胞增殖与阿霉素耐药  

GSTP1 promote breast cancer cell proliferation and adriamycin resistance by regulating STAT3 signal pathway

作　　者：戴素华 嵇晓艳[1] 戴夕超 DAI Suhua;JI Xiaoyan;DAI Xichao(Department of Oncology,Bin Hai County People′s Hospital,Yancheng 224500,China)

机构地区：[1]滨海县人民医院肿瘤科,江苏盐城224500 [2]盐城市第一人民医院肿瘤科,224000

出　　处：《临床肿瘤学杂志》2024年第2期114-119,共6页Chinese Clinical Oncology

基　　金：江苏省卫生健康委课题(Z2021041);盐城市课题(YK2018017);盐城市第一人民医院院课题重点项目(ZD2018004)。

摘　　要：目的 探讨谷胱甘肽S-转移酶P1(GSTP1)对人乳腺癌MCF-7细胞增殖与阿霉素耐药性的影响及其作用机制。方法 Western blotting检测野生型乳腺癌细胞MCF-7和阿霉素耐药乳腺癌细胞MCF-7/ADR中的GSTP1表达量,通过在MCF-7细胞中转染Flag-GSTP1质粒过表达GSTP1,在MCF-7/ADR细胞中转染GSTP1敲低慢病毒(shGSTP1)干扰GSTP1表达;平板克隆形成实验、CCK-8法和流式细胞术检测转染后乳腺癌细胞的增殖能力、阿霉素耐药性及凋亡水平的变化。Western blotting检测GSTP1表达水平的变化对STAT3通路激活的影响。结果 GSTP1在MCF-7细胞中表达量极低,且显著低于MCF-7/ADR细胞系。GSTP1过表达的MCF-7/ADR细胞克隆形成数量显著多于野生型MCF-7细胞(P<0.05)。过表达GSTP1显著提升了MCF-7细胞的增殖能力,而在MCF-7/ADR细胞系干扰GSTP1表达水平后显著抑制了细胞增殖能力(P<0.05)。CCK-8结果显示,在0.1、1、10、50μmol/ml不同浓度的阿霉素处理下,GSTP1表达水平与MCF-7细胞对阿霉素的耐药性呈正相关(P<0.05)。流式细胞术检测结果显示,GSTP1过表达组(Flag-GSTP1)和对照组(Flag)的凋亡率分别为(11.41±1.16)%和(21.1±1.72)%,GSTP1过表达显著抑制了阿霉素诱导的细胞凋亡(P<0.05)。Western blotting检测结果显示,GSTP1的过表达激活了STAT3信号通路,同时在MCF-7/ADR细胞系中抑制STAT3显著降低了GSTP1的表达水平(P<0.05)。结论 GSTP1通过上调STAT3表达调节乳腺癌细胞MCF-7的增殖能力和对阿霉素的耐药性。Objective To investigate the effect and mechanism of glutathione S-transferase P1(GSTP1)on proliferation and adriamycin resistance of breast cancer MCF-7 cells.Methods The expression levels of GSTP1 in MCF-7 and MCF-7/ADR cell lines were assessed via Western blotting.Overexpression of GSTP1 was achieved by transfection of Flag-GSTP1 plasmid in MCF-7 cells,and interference with GSTP1 expression was achieved by transfection of GSTP1 knockdown lentivirus(shGSTP1)in MCF-7/ADR cells.Proliferation,doxorubicin resistance and apoptosis levels of the cell lines after transfection were evaluated via colony formation assay,CCK-8 method and flow cytometry.Finally,the impact of GSTP1 expression level on the activation of the STAT3 pathway was investigated through Western blotting.Results GSTP1 exhibited markedly low expression in MCF-7 cells,significantly lower than that in the MCF-7/ADR cell line.The number of clone formation in MCF-7/ADR cells overexpressing GSTP1 was significantly higher than that in wild-type MCF-7 cells(P<0.05).Overexpression of GSTP1 significantly enhanced the proliferative capacity of MCF-7 cells,whereas interference with GSTP1 expression levels in the MCF-7/ADR cell line significantly inhibited cell proliferation(P<0.05).CCK-8 result demonstrated a positive correlation between GSTP1 expression levels and doxorubicin resistance in MCF-7 cells treated with doxorubicin concentrations of 0.1,1,10,and 50μmol/ml(P<0.05).Flow cytometry analysis revealed that the apoptosis rates of the GSTP1 overexpression group(Flag-GSTP1)and the control group(Flag)were(11.41±1.16)%and(21.1±1.72)%,respectively.GSTP1 overexpression significantly inhibited doxorubicin-induced cell apoptosis(P<0.05).Western blotting further confirmed that high expression of GSTP1 activated the STAT3 signaling pathway,while inhibiting STAT3 in the MCF-7/ADR cell line significantly reduced the expression levels of GSTP1(P<0.05).Conclusion GSTP1 regulates the proliferation of MCF-7 cells and drug resistance to adriamycin by up-regulating STAT3

关 键 词：乳腺癌 谷胱甘肽S转移酶P1 增殖 阿霉素耐药 

分 类 号：R737.9[医药卫生—肿瘤]