摘要
目的探究趋化因子受体7(CXCR7)沉默对人肾癌细胞增殖、侵袭及凋亡的影响。方法采用Western blot法筛选CXCR7高表达的肾癌细胞系,检验慢病毒转染对CXCR7蛋白的沉默效果;沉默CXCR7蛋白的表达后,利用WST-8细胞增殖实验及细胞生长曲线检测肾癌细胞的增殖能力;采用Transwell实验、划痕实验检测肾癌细胞的侵袭和迁移能力;采用流式细胞仪检测无血清诱导肾癌细胞的凋亡情况;采用Western blot法检测肾癌细胞中B细胞淋巴瘤/白血病-2(Bcl-2)、Bax、cleaved-caspase-3、血管内皮生长因子(VEGF)、基质金属蛋白酶(MMP)2、MMP9蛋白的表达水平。结果 CXCR7蛋白在肾癌组织中的表达水平高于健康肾组织(P﹤0.05)。Western blot检测结果显示,CXCR7蛋白在786-O肾癌细胞中的表达水平相对较高,选择其用于后续研究CXCR7蛋白对肾癌细胞的影响。慢病毒转染沉默786-O细胞中CXCR7蛋白的表达后,786-O细胞的增殖、侵袭和迁移能力均低于对照组和sh-NC组;无血清诱导后,786-O细胞的凋亡率升高及Bax、cleaved-caspase-3的表达水平均升高,Bcl-2、VEGF、MMP2、MMP9的表达水平均降低(P﹤0.05)。结论 CXCR7通过调控凋亡相关蛋白的表达抑制凋亡信号,促进肿瘤新生血管形成,从而促进人肾癌细胞的增殖和侵袭。
Objective To explore the effects of silencing CXC chemokine receptor 7(CXCR7)on proliferation,invasion and apoptosis of human renal carcinoma cells.Method The renal carcinoma cell lines with high CXCR7 expression were screened by Western blot.The silencing effects of lentiviral transfection on CXCR7 protein were tested.After silencing the expression of cell CXCR7 protein,the proliferation ability of renal carcinoma cells was detected by WST-8 cells proliferation experiment and cell growth curves.The invasion and migration abilities of renal carcinoma cells were detected by Transwell assay and scratch wound assay.Apoptosis of renal carcinoma cells with serum-free induction was detected by flow cytometry.The expression levels of B cell lymphoma/leukemia-2(Bcl-2),Bax,cleaved-caspase-3,vascular endothelial growth factor(VEGF),MMP2 and MMP9 in renal carcinoma cells were detected by Western blot.Result The expression level of CXCR7 protein in renal carcinoma cells was higher than that in healthy renal tissues(P<0.05).The test results of Western blot showed that expression level of CXCR7 protein was relatively higher in renal carcinoma 786-O cells,which were selected for subsequent study on the effects of CXCR7 protein on renal carcinoma cells.After lentivirus transfection that silenced the expression of CXCR7 protein in 786-O cells,the proliferation,invasion and migration abilities of 786-O cells were lower than those in control group and sh-NC group.After serum-free induction,apoptosis rate of 786-O cells was increased,expression levels of Bax and cleared-caspase-3 were increased,and expression levels of Bcl-2,VEGF,MMP2 and MMP9 were decreased(P<0.05).Conclusion CXCR7 inhibits apoptosis signals,promotes the formation of tumor neovascularization,and thus enhances the proliferation and invasion abilities of human renal carcinoma cells by regulating the expression of apoptosis-related proteins.
作者
吕栋
滕志刚
周云飞
陈芙蓉
LYU Dong;TENG Zhigang;ZHOU Yunfei;CHEN Furong(Department of Urologic Surgery,Kaifeng Central Hospital,Kaifeng 475000,He’nan,China;Department of Pathology,Kaifeng Central Hospital,Kaifeng 475000,He’nan,China)
出处
《癌症进展》
2020年第10期1043-1046,1079,共5页
Oncology Progress
关键词
趋化因子受体7
基因沉默
凋亡
肾癌
增殖
侵袭
CXC chemokine receptor 7
gene silencing
apoptosis
renal carcinoma
proliferation
invasion
