下调ClC-5表达抑制多发性骨髓瘤细胞增殖并诱导凋亡  

Down-regulation of ClC-5 expression inhibits proliferation and induces apoptosis in multiple myeloma cells

作　　者：苗玉迪[1] 胡星星 李艳春 Miao Yudi;Hu Xingxing;Li Yanchun(Department of Hematopathology,Shaanxi Provincial People’s Hospital,Xi’an 710068,China;Laboratory Diagnostic Center,Xi‘an International Medical Center Hospital Blood Disease Hospital,Xi’an 710018,China)

机构地区：[1]陕西省人民医院血液内科,西安710068 [2]西安国际医学中心医院血液病医院实验诊断中心,西安710018

出　　处：《中国组织化学与细胞化学杂志》2022年第3期239-245,266,共8页Chinese Journal of Histochemistry and Cytochemistry

基　　金：陕西省自然科学基础研究计划(2021JM-548)。

摘　　要：目的探讨氯通道5(chloride channel 5,ClC-5)对多发性骨髓瘤(multiple myeloma,MM)细胞凋亡的作用及其机制。方法用免疫荧光和Western blot法检测MM细胞系(ARH77、U266和RPMI8266)和外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中ClC-5的表达。ClC-5 siRNA转染MM细胞和PBMC细胞后,CCK-8法检测细胞活性。敲降ARH77和U266细胞中ClC-5后,用Hoechst 33258染色观察细胞核形态,Annexin V-FITC/PI双染色流式细胞仪检测细胞凋亡,JC-1染色检测线粒体膜电位,Western blot检测细胞色素c(cytochrome c,cyt-c)释放和Cleaved Caspase-3表达。结果与PBMC细胞比较,ClC-5在MM细胞中表达升高。敲降ClC-5后,ARH77和U266的细胞活性降低,但PBMC的细胞活性不变。进一步研究显示,敲降ClC-5能导致ARH77和U266的细胞核固缩,凋亡增加,线粒体膜电位降低,促进线粒体cyt-c释放到细胞质,并增加Cleaved Caspase-3表达。结论敲降ClC-5能通过线粒体凋亡通路来促进MM细胞凋亡。Objective To investigate the effect and mechanism of chloride channel 5(ClC-5)on apoptosis of multiple myeloma(MM)cells.Methods The expression of ClC-5 in MM cell lines(ARH77,U266 and RPMI8266)and peripheral blood mononuclear cells(PBMC)was determined by immunofluorescence and Western blot.MM cells and PBMC cells were transfected with ClC-5 siRNA,and the cell viability was detected by CCK-8 assay.After knockdown of ClC-5 in ARH77 and U266 cells,the nuclear morphology was observed by Hoechst 33258 staining,apoptosis was detected by flow cytometry with Annexin V-FITC/PI double staining,mitochondrial membrane potential was detected by JC-1 staining,cytochrome c(cyt-c)release and Cleaved caspase-3 expression were detected by Western blot.Results Compared with PBMC cells,the expression of ClC-5 was increased in MM cells.After knockdown of ClC-5,the cell viability of ARH77 and U266 was decreased,but the cell viability of PBMC remained unchanged.Further studies showed that knockdown ClC-5 could lead to nuclear pyrosis of ARH77 and U266,increase apoptosis,decrease mitochondrial membrane potential,promote the release of mitochondrial cyt-c into the cytoplasm,and increase Cleaved caspase-3 expression.Conclusion Knockdown of ClC-5 can promote MM cell apoptosis through mitochondrial apoptotic pathway.

关 键 词：多发性骨髓瘤 氯通道5 凋亡 线粒体凋亡通路 

分 类 号：R733.3[医药卫生—肿瘤]