摘要 目的探讨跨膜蛋白16A(TMEM16A)在乳腺癌细胞中参与信号通路的富集分析及其对乳腺癌患者预后的影响,探究其对雌激素受体(ER)和孕酮受体(PR)阳性乳腺癌促进增殖作用的机制。方法从公共数据库癌症基因组图谱在线数据库中下载乳腺癌的转录组及临床数据,根据TMEM16A表达的中位值将患者分为高表达组和低表达组,用基因集富集分析进行基因富集和通路分析。用Kaplan-Meier法进行生存分析。用Western Blot方法检测过表达及敲减TMEM16A后ER和PR阳性乳腺癌T47D细胞的TMEM16A蛋白及下游信号通路磷酸化表皮生长因子受体(p-EGFR)和EGFR蛋白的表达情况。用CCK-8细胞增殖实验检测乳腺癌T47D细胞转染过表达TMEM16A质粒及用EGFR通路抑制剂后的细胞增殖能力。结果高表达TMEM16A组乳腺癌患者的基因高度富集于EGFR通路、葡萄糖激酶调节蛋白通路和间质表皮转化因子(MET)等通路。Kaplan-Meier生存分析显示:TMEM16A高表达乳腺癌患者预后生存期显著降低,TMEM16A与EGFR共同高表达组的总生存显著低于共同低表达组。Western Blot结果显示:过表达TMEM16A能够增加p-EGFR蛋白表达,反之敲减TMEM16A后p-EGFR蛋白表达水平降低。CCK-8结果显示:转染空质粒与过表达(TMEM16A OE)的相对增殖力分别为(100.00±0)%和(121.40±5.99)%,并且这种促增殖作用可以被吉非替尼所逆转,TMEM16A OE+吉非替尼为(78.26±4.74)%。结论 TMEM16A可能通过EGFR信号通路促进ER、PR阳性乳腺癌T47D细胞的增殖。 Objective To explore the enrichment analysis of transmembrane protein 16 A(TMEM16 A)-mediated signaling pathways in breast cancer cells and its effect on the prognosis of breast cancer patients,and explore its mechanism of promoting the proliferation of estrogen receptor(ER)and progesterone receptor(PR)positive breast cancer.Methods The transcriptome and clinical data of breast cancer were downloaded from the public The Cancer Genome Atlas database.According to the median value of TMEM16 A expression,patients were divided into high and low expression groups,and gene set enrichment analysis(GSEA)was used for gene enrichment analysis and pathway analysis.Kaplan-Meier was used for survival analysis.Western Blot was used to detect the expressions of TMEM16 A,phosphorylated epidermal growth factor receptor(p-EGFR),EGFR in T47 D breast cancer cells after overexpression and knockdown of TMEM16 A.CCK-8 cell proliferation test was used to detect the cell proliferation ability of TMEM16 A-overexpressing T47 D breast cancer cells in the presence of EGFR inhibitors.Results The genes of breast cancer patients with high expression of TMEM16 A were highly enriched in EGFR pathway,glucokinase regulatory protein pathway and mesenchymal to epithelial transition factor(MET)pathway et al.Kaplan-Meier survival analysis showed that high TMEM16 A expression was associated with poor prognosis of breast cancer patients,and high TMEM16 A/EGFR expression was associated with shorter overall survival in breast cancer patients compared with low TMEM16 A/EGFR expression.Western Blot results showed TMEM16 A overexpression increased,and TMEM16 A knockdown decreased phosphorylated EGFR expression in T47 D breast cancer cells.CCK-8 results showed that the relative proliferation of transfected empty plasmid and overexpression(TMEM16 A OE)was(100.00±0)%and(121.40±5.99)%,and this proliferation effect could be reversed by gefitinib,the relative proliferation of TMEM16 A OE+gefitinib was(78.26±4.74)%.Conclusion TMEM16 A may promote the proliferation
机构地区 中国医科大学药学院离子通道药理学研究室
出处 《中国临床药理学杂志》 CAS CSCD 北大核心 2021年第7期832-836,共5页 The Chinese Journal of Clinical Pharmacology
基金 国家自然科学基金资助项目(81572613)。
关键词 跨膜蛋白16A 乳腺癌 基因集富集分析 生存分析 表皮生长因子受体 transmembrane protein 16A breast cancer gene set enrichment analysis survival analysis epidermal growth factor receptor
分类号 R97 [医药卫生—药品]
