ETF在乳腺癌组织中的表达及对乳腺癌细胞增殖的影响  

Expression of ETF in breast cancer tissue and its effect on the proliferation of breast cancer cells

作　　者：赵璐 孙茂伟 邹鹏 ZHAO Lu;SUN Maowei;ZOU Peng(Department of Biochemistry and Molecular Biology,Shenyang Medical College,Shenyang 110034,China;Department of General Surgery,Jishuitan Hospital of Shenyang)

机构地区：[1]沈阳医学院基础医学院生物化学与分子生物学教研室,辽宁沈阳110034 [2]沈阳市积水潭医院普外科

出　　处：《沈阳医学院学报》2021年第5期421-426,共6页Journal of Shenyang Medical College

基　　金：国家自然科学基金(No.81270574)。

摘　　要：目的:探究ETF在乳腺癌组织中的表达以及对乳腺癌细胞增殖的影响。方法:收集沈阳市积水潭医院普外科2018年1月至2020年12月经病理组织学确诊的76例乳腺癌患者癌组织和癌旁组织,采用免疫组化技术、实时荧光定量PCR(RT-qPCR)、Western blot法检测ETF在乳腺癌及癌旁组织中的表达,同时检测其在人正常乳腺细胞MCF10A、人乳腺癌细胞MDA-MB-231和MCF-7中的表达。通过慢病毒载体构建过表达ETF的细胞株MDA-MB-231-ETF/MCF-7-ETF和其阴性对照组稳转细胞株MDA-MB-231-NC/MCF-7-NC,采用CCK-8实验检测细胞增殖;Annexin V-FITC/PI染色检测细胞周期,RT-qPCR、Western blot检测周期相关蛋白表达情况。结果:ETF蛋白在乳腺癌组织中阳性表达率为73.68%(56/76),高于在癌旁组织中阳性表达率[17.11%(13/76)](P<0.01);乳腺癌组织中ETF mRNA和蛋白表达水平显著高于癌旁正常组织(P<0.01)。MDA-MB-231和MCF-7细胞中ETF mRNA和蛋白表达水平显著高于MCF10A细胞(P<0.01)。与阴性对照组和空白对照组比较,MDA-MB-231-ETF/MCF-7-ETF组细胞在48、72、96 h处细胞增殖显著提高(P<0.01);细胞周期中G_(2)/M期细胞比例显著上升,细胞周期进程加快(P<0.01)。MDA-MB-231-ETF/MCF-7-ETF组Cyclin D1、CDK4、CDK6、Cyclin E1 mRNA和蛋白表达均显著高于阴性对照组和空白对照组(P<0.01),RB mRNA和蛋白水平与阴性对照组与空白对照组比较差异无统计学意义(P>0.05),其磷酸化水平显著高于阴性对照组和空白对照组(P<0.01)。结论:ETF在乳腺癌组织和细胞中高表达,通过调控细胞周期和相关蛋白表达促进乳腺癌细胞增殖。Objective:To investigate the expression of ETF in breast cancer and its effect on the proliferation of breast cancer cells.Methods:Breast cancer tissues and adjacent tissues from 76 patients with breast cancer diagnosed by pathology were collected from department of general surgery,Jishuitan Hospital from Jan 2018 to Dec 2020.The expression of ETF in breast cancer tissues and adjacent tissues were detected by immunohistochemistry,Western blot and RT-qPCR.Meanwhile,the expression of ETF in human normal breast cells MCF10A,human breast cancer MDA-MB-231 and MCF-7 cells were measured.Further,MDA-MB-231-ETF and MCF-7-ETF overexpressing ETF and its negative control group were constructed by lentiviral vector.And then the CCK-8,Annexin V-FITC/PI double staining flow cytometry,Western blot and RT-qPCR were used to determine cell proliferation activity,cell cycle and the expression of cell cycle related proteins,respectively.Results:The positive expression rate of ETF protein was 73.68%(56/76)in breast cancer tissues and was higher than that in adjacent tissues which was 17.11%(13/76)(P<0.01).The expression levels of ETF mRNA and protein in breast cancer tissues were significantly higher than those in adjacent tissues(P<0.01).The expression levels of ETF mRNA and protein in MDA-MB-231/MCF-7 cells were significantly higher than those in MCF10A cells(P<0.01).The cell proliferation and the proportion of G_(2)/M phase cells in MDA-MB-231-ETF/MCF-7-ETF group were significantly higher than those in negative control and blank control groups(P<0.01).Moreover,the mRNA and protein expressions of Cyclin D1,CDK4,CDK6,and Cyclin E1 in the MDA-MB-231-ETF/MCF-7-ETF group were significantly higher than those in negative control and blank control groups(P<0.01).There was no significant difference in the mRNA and protein expression of RB compared to negative control and blank control group(P>0.05),while the level of p-RB was significantly higher than that in negative control and blank control groups(P<0.01).Conclusion:The expression level

关 键 词：乳腺癌 ETF 增殖 周期 

分 类 号：R737.9[医药卫生—肿瘤]