ROR1-AS1调控Notch1信号通路对乳腺癌MCF-7细胞增殖和凋亡影响  

Effect of ROR1-AS1 Regulating Notch1 Signaling Pathway on Proliferation and Apoptosis of Breast Cancer MCF-7 Cells

作　　者：韩拓 肖栋[1] 陈亮[1] 张伟[2] HAN Tuo;XIAO Dong;CHEN Liang;ZHANG Wei(3201 Hospital Affiliated to Xi'an Jiaotong University College of Medicine,Hanzhong 723000,China;The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China)

机构地区：[1]西安交通大学医学院附属三二〇一医院,陕西汉中723000 [2]西安交通大学附属第一医院,陕西西安710061

出　　处：《肿瘤学杂志》2022年第1期40-45,共6页Journal of Chinese Oncology

基　　金：陕西省自然科学基础研究计划[一般项目(面上)(2020JM-393)]。

摘　　要：[目的]研究酪氨酸蛋白激酶跨膜受体1反义RNA 1(tyrosine protein kinase transmembrane receptor 1 antisense RNA 1,ROR1-AS1)调控Notch1信号通路对乳腺癌MCF-7细胞增殖、凋亡及上皮-间质转化(epithelial-mesenchymal transition,EMT)的影响。[方法]实时荧光定量PCR方法检测乳腺癌MCF-7、MDAMB-231、MDA-MB-361细胞和正常乳腺上皮MCF10A细胞中ROR1-AS1表达变化。乳腺癌MCF-7细胞分成对照、sh-NC(转染shRNA对照)、sh-ROR1-AS1(转染ROR1-AS1 shRNA)、sh-ROR1-AS1+Jagged1组(转染ROR1-AS1 shRNA,Notch1通路激活剂Jagged1处理)。CCK-8实验检测细胞增殖,流式细胞术检测细胞凋亡,Transwell小室检测细胞迁移和侵袭,Western blot方法检测细胞中活化的半胱氨酰天冬氨酸特异性蛋白酶9(cleaved cysteinyl aspartate-specific protease-9,C-Caspase-9)、上皮型钙黏蛋白(E-cadherin)、活化的半胱氨酰天冬氨酸特异性蛋白酶3(cleaved cysteinyl aspartate-specific protease-3,C-Caspase-3)、神经型钙黏蛋白(N-cadherin)、Notch1、Hes1蛋白表达。[结果]乳腺癌MCF-7、MDA-MB-231、MDA-MB-361细胞中ROR1-AS1水平明显高于正常乳腺上皮MCF10A细胞(P<0.05)。与对照组、sh-NC组比较,sh-ROR1-AS1组乳腺癌MCF-7细胞存活率降低(100.00%±9.82%,99.74%±12.05%,58.91%±6.33%),细胞凋亡率升高(4.81%±0.26%,4.93%±0.34%,20.58%±2.11%),细胞迁移数目(225.36±12.84个,224.89±19.56个,140.89±13.64个)和侵袭数目(180.47±16.32个,177.54±16.92个,110.44±10.87个)减少,细胞中C-Caspase-9、E-cadherin、C-Caspase-3蛋白表达水平增加,N-cadherin、Notch1、Hes1蛋白表达水平降低(P<0.05)。与sh-ROR1-AS1组比较,shROR1-AS1+Jagged1组乳腺癌MCF-7细胞存活率升高(100.00%±11.58%vs 147.36%±12.05%),细胞凋亡率降低(19.54%±1.74%vs 8.36%±0.75%),细胞迁移数目(139.58±12.78个vs 175.26±16.94个)和侵袭数目(107.45±9.35个vs 162.04±13.67个)均升高,C-Caspase-9、E-cadherin、C-Caspase-3蛋白水平降低,N-cadherin、Notch1、Hes1蛋白水平升高(P<0.05)[Objective]To investigate the effect of long non-coding RNA tyrosine protein kinase transmembrane receptor 1 antisense RNA 1(lncRNA ROR1-AS1)regulating Notch1 signaling pathway on the proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of breast cancer cells.[Methods]The expression of ROR1-AS1 in human breast cancer MCF-7,MDA-MB-231,MDA-MB-361 cells and normal breast epithelial MCF10 A cells were detected by real-time PCR method.Breast cancer MCF-7 cells were transfected with shRNA-NC(shNC group),ROR1-AS1 shRNA(sh-ROR1-AS1 group)or ROR1-AS1 shRNA and Notch1 pathway activator Jagged1(sh-ROR1-AS1+Jagged1 group).Cell proliferation was detected with CCK-8 method,cell apoptosis was detected with flow cytometry,cell migration and invasion was detected with transwell assay,the expression of C-Caspase-9,E-cadherin,C-Caspase-3,N-cadherin,Notch1 and Hes1 proteins was detected with Western blot.[Results]The expression levels of ROR1-AS1 in breast cancer MCF-7,MDA-MB-231 and MDA-MB-361 cells were significantly higher than that in normal breast epithelial MCF10 A cells(P<0.05).Compared with the control and sh-NC groups,in sh-ROR1-AS1 group the cell survival rate decreased(100.00%±9.82%,99.74%±12.05%,58.91%±6.33%),cell apoptosis rate increased(4.81%±0.26%,4.93%±0.34%,20.58%±2.11%),cell migration(225.36±12.84,224.89±19.56,140.89±13.64)and invasion number(180.47±16.32,177.54±6.92,110.44±10.87)decreased,the expression levels of C-Caspase-9,E-cadherin and C-Caspase-3 proteins increased,the expression levels of N-cadherin,Notch1,Hes1 proteins decreased(P<0.05).Compared with the shROR1-AS1 group,in the sh-ROR1-AS1+Jagged1 group,the survival rate increased(100.00%±11.58%vs147.36%±12.05%),cell apoptosis rate(19.54%±1.74%vs 8.36%±0.75%)decreased,cell migration(139.58±12.78 vs 175.26±16.94)and invasion numbers(107.45±9.35 vs 162.04±13.67)increased,the expression levels of C-Caspase-9,E-cadherin,C-Caspase-3 proteins decreased,and the expression levels of N-cadherin,Notch1,Hes1 proteins increased(P<0.05).

关 键 词：ROR1-AS1 乳腺癌 上皮-间质转化 凋亡 Notch1信号通路 

分 类 号：R737.9[医药卫生—肿瘤]