CXCR2在膀胱癌干细胞增殖、凋亡及侵袭转移中的作用及机制  

Role of CXCR2 in proliferation,invasion,and metastasis of bladder cancer stem cells

作　　者：李顺来[1] 姜亭起[1] 马天加[2] LI Shunlai;JIANG Tingqi;MA Tianjia(Jinan Fifth People's Hospital,Jinan 250022,China)

机构地区：[1]济南市第五人民医院,济南250022 [2]山东大学第二医院

出　　处：《山东医药》2020年第14期17-20,共4页Shandong Medical Journal

基　　金：山东省医药卫生科技发展计划项目(2017WS668)。

摘　　要：目的探讨趋化因子受体2(CXCR2)在膀胱癌干细胞增殖、凋亡及侵袭转移中的作用及机制。方法取膀胱癌肿瘤组织,培养成悬浮肿瘤细胞球,用流式细胞仪筛选出CD44+膀胱癌干细胞。采用Western blotting法检测干细胞标志基因Oct4、Sox2蛋白表达,并检测CXCR2蛋白表达。培养膀胱癌干细胞,将其分为sh-CXCR2组和转染对照组,sh-CXCR2组加入慢病毒转染试剂polybrene与CXCR2敲减慢病毒,转染对照组加入慢病毒转染试剂polybrene与对照scramble。转染72 h后,采用MTS法检测两组细胞增殖率,软琼脂克隆实验观察细胞克隆形成能力,流式细胞仪检测细胞凋亡率,Transwell小室转移实验和Boyden侵袭实验观察细胞转移和侵袭能力,Western blotting法检测两组细胞中凋亡相关蛋白FOXO1A、Bcl-2以及侵袭转移相关蛋白MMP-2、MMP-9的相对表达量。结果流式细胞仪分选出CD44+细胞占全部细胞的2.37%,CD44+细胞中Oct4、Sox2、CXCR2蛋白表达水平高于CD44-细胞(P均<0.05)。与转染对照组比较,sh-CXCR2组细胞增殖率降低,克隆球形成数量减少,细胞凋亡率升高,侵袭和转移实验的穿膜细胞数量减少,细胞中FOXO1A、Bcl-2、MMP-2、MMP-9蛋白表达降低(P均<0.05)。结论CXCR2在膀胱癌干细胞中高表达,抑制其表达可以降低膀胱癌干细胞的增殖、侵袭、转移能力,促进细胞凋亡;其机制可能是通过作用于凋亡相关蛋白FOXO1A、Bcl-2和侵袭转移相关蛋白MMP-2、MMP-9实现的。Objective To investigate the role and mechanism of chemokine receptor 2(CXCR2)in the proliferation,invasion,and metastasis of bladder cancer stem cells.Methods The tumor tissues of bladder cancer were collected and were cultured into suspension tumor cell spheres.CD44+bladder cancer stem cells were screened out by flow cytometry.Western blotting was used to detect the expression of stem cell marker genes Oct4 and Sox2,and CXCR2 protein was detected.The bladder cancer stem cells were cultured and then were divided into the sh-CXCR2 group and sh-scramble group.We added lentiviral transfection reagent polybrene and CXCR2 knockdown lentivirus in the sh-CXCR2 group,and added lentiviral transfection reagent polybrene and control scramble to the sh-scramble group.After 72 hours of transfection,the cell proliferation ability,tumor sphere formation ability,apoptosis rate,and invasion and metastasis abilities of the two groups were measured by MTS test,soft agar cloning test,flow cytometry,Transwell chamber transfer test,and Boyden invasion test.Western blotting was used to detect the expression of apoptosis-related proteins FOXO1A,Bcl-2,and invasion and metastasis-related proteins matrix metalloproteinase 2(MMP-2)and MMP-9.Results CD44+cells sorted by flow cytometry accounted for 2.37%of the total cells.The expression levels of Oct4,Sox2,and CXCR2 proteins in the CD44+cells were significantly higher than those of CD44-cells.Compared with the sh-scramble group,the proliferation ability of bladder cancer stem cells decreased,the tumor ball formation ability decreased,the cell apoptosis rate increased,and the invasion and metastasis abilities decreased,and the FOXO1A,Bcl-2,MMP-2,and MMP-9 expression levels decreased in the bladder cancer stem cells of the sh-CXCR2 group(all P<0.05).Conclusion CXCR2 is highly expressed in bladder cancer stem cells,and inhibiting its expression can reduce the proliferation,invasion,and metastasis of bladder cancer stem cells,and promote apoptosis by acting on apoptosis-related proteins FOXO1A,

关 键 词：膀胱癌干细胞 趋化因子受体2 细胞增殖 细胞凋亡 肿瘤侵袭 肿瘤转移 

分 类 号：R737.21[医药卫生—肿瘤]