IL-24对乳腺癌细胞CXCR4表达及肿瘤细胞侵袭、迁移及凋亡能力的影响  

Effects of IL-24 on CXCR4 expression and invasion,migration and apoptosis of breast cancer cell

作　　者：蒋雅玲 胡学丽 陈辉 段智 刘景 范先成 JIANG Yaling;HU Xueli;CHEN Hui;DUAN Zhi;LIU Jing;FAN Xiancheng(Department of Breast-Nail Surgery,Changsha First Hospital,Changsha 410005,China)

机构地区：[1]长沙市第一医院乳甲外科,长沙410005 [2]长沙市第一医院病理科,长沙410005

出　　处：《中国免疫学杂志》2022年第16期1977-1982,共6页Chinese Journal of Immunology

基　　金：湖南省卫生健康委科研计划项目(20200301)资助。

摘　　要：目的:探究过表达IL-24对乳腺癌细胞CXCR4表达及肿瘤细胞侵袭、迁移及凋亡的影响与机制。方法:免疫组织化学染色(IHC)检测乳腺癌与癌旁正常乳腺组织CXCR4与其配体CXCL12表达,并分析其相关性;Western blot检测CXCR4在人正常乳腺上皮细胞MCF-10A及乳腺癌细胞系MCF-7、MDA-MB-453和MDA-MB-231中的表达;采用慢病毒稳转技术在乳腺癌细胞系MDA-MB-231中过表达IL-24,免疫荧光显微镜、RT-PCR和ELISA检测感染效率;将细胞分为5组:LV-IL-24组(感染过表达IL-24慢病毒)、LV-NC组(感染阴性对照慢病毒)、AMD3100组(采用CXCR4/CXCL12轴阻滞剂AMD3100进行培养)、LV-IL-24+AMD3100组(采用AMD3100进行培养的感染过表达IL-24慢病毒的细胞)、阴性对照组(正常培养的MDAMB-231细胞);Transwell、划痕实验和Annexin V-FITC细胞凋亡实验检测上述各组细胞侵袭、迁移及凋亡;Western blot检测各组细胞CXCR4、AKT与mTOR蛋白磷酸化水平及凋亡相关蛋白Bax与cleaved-Caspase-3表达。结果:CXCR4与CXCL12在乳腺癌组织中的表达均显著高于正常乳腺组织,且二者表达呈正相关;与MCF-10A细胞相比,CXCR4在乳腺癌细胞系MCF-7、MDA-MB-453及MDA-MB-231中表达明显增加(P<0.05),以MDA-MB-231最为显著(P<0.01)。转染过表达IL-24的慢病毒后,MDA-MB-231细胞IL-24 mRNA与蛋白表达均显著增加(P<0.05);与阴性对照组相比,LV-IL-24组、AMD3100组和LV-IL-24+AMD3100组细胞侵袭、迁移能力均显著降低(P<0.05),而凋亡率明显提高(P<0.05),LV-IL-24+AMD3100组变化最为显著(P<0.01);Western blot结果显示,与阴性对照组相比,LV-IL-24组、AMD3100组和LV-IL-24+AMD3100组细胞CXCR4、AKT与mTOR蛋白磷酸化水平显著降低(P<0.05),而凋亡相关蛋白Bax与cleaved-Caspase-3表达明显增加(P<0.05),LV-IL-24+AMD3100组变化最为显著(P<0.01)。结论:乳腺癌细胞中过表达IL-24可抑制CXCR4表达,通过下调PI3K/AKT/mTOR信号通路降低肿瘤细胞侵袭、迁移能力,促进其凋亡,联合使用Objective:To explore effect of IL-24 overexpression on CXCR4 expression,invasion,migration and apoptosis of breast cancer cells and its mechanism.Methods:Immunohistochemical staining(IHC)was used to detect expressions of CXCR4 and its ligand CXCL12 in breast cancer and paracancerous normal breast tissues,whose correlation was further analyzed;Western blot was used to detect expression of CXCR4 in normal human mammary epithelial cells MCF-10A and breast cancer cell lines MCF-7,MDA-MB-453 and MDA-MB-231. IL-24 was overexpressed in breast cancer cell line MDA-MB-231 by lentivirus stabilization technique,whose infection efficiency was detected by immunofluorescence microscopy,RT-PCR and ELISA. Cells were divided into5 groups:LV-IL-24 group(infected with IL-24 lentivirus),LV-NC group(infected with negative control lentivirus),AMD3100 group(cultured with CXCR4/CXCL12 axis blocker AMD3100),LV-IL-24+AMD100 group(infected with IL-24 lentivirus and cultured with AMD3100),negative control group(MDA-MB-231 cells in normal culture). Transwell,scratch test and Annexin V-FITC cell apoptosis test were used to detect invasion,migration and apoptosis of cells;Western blot was used to detect phosphorylation levels of CXCR4,AKT and mTOR proteins,and expressions of Bax and cleaved-Caspase-3 proteins.Results:Expressions of CXCR4 and CXCL12 in breast cancer tissues were significantly higher than those in normal breast tissue,and expressions of CXCR4 and CXCL12were positively correlated. Compared with MCF-10A cells,expressions of CXCR4 in breast cancer cell lines MCF-7,MDA-MB-453and MDA-MB-231 were significantly increased(P<0.05),and MDA-MB-231 was the most significant(P<0.01). mRNA and protein levels of IL-24 in MDA-MB-231 cells were significantly increased after infected with IL-24 lentivirus(P<0.05). Compared with control group,invasion and migration abilities of tumor cells in LV-IL-24 group,AMD3100 group and LV-IL-24+AMD3100 group were significantly decreased(P<0.05),while apoptosis rate was significantly increased(P<0.05),especial

关 键 词：乳腺癌 IL-24 CX-C趋化因子受体4 慢病毒感染 

分 类 号：R737.9[医药卫生—肿瘤]