CMTM5表达对多发性骨髓瘤细胞增殖的影响及其机制研究  

Pathophysiologic mechanism of CMTM5 low expression in multiple myeloma progression

作　　者：马媛 史进方[1] 仇惠英 袁靖 章扬 周鹏[1] 徐晶晶[1] 韩清珍[1] Ma Yuan;Shi Jinfang;Qiu Huiying;Yuan Jing;Zhang Yan;Zhou Peng;Xu Jingjing;Han Qingzhen(Center for Clinical Labrotary,the First Affiliated Hospital of Soochow University,Suzhou 215006,China;Institute of Hematology of Jiangsu,Suzhou 215006,China;Clinical Laboratory,the Second Affiliated Hospital of Anhui Medical Univercity,Hefei 230001,China)

机构地区：[1]苏州大学附属第一医院临床检测中心,215006 [2]江苏血液研究所,苏州215006 [3]安徽医科大学附属第二医院检验科,合肥230001

出　　处：《中华血液学杂志》2019年第1期58-62,共5页Chinese Journal of Hematology

基　　金：国家自然科学基金青年基金(81501425);江苏省医学创新团队(CXTDB2017009).

摘　　要：目的探索趋化素样因子超家族成员CMTM5表达对多发性骨髓瘤(MM)细胞增殖活性的影响及其机制。方法去甲基化药物地西他滨处理MM细胞系U266细胞,采用实时荧光定量PCR法检测处理前后U266细胞CMTM5、caspase3、caspase9的表达水平并分析其相关性;pcDNA3.1质粒转染U266细胞使CMTM5过表达后,采用CCK-8法检测细胞增殖活性变化。结果①与对照组相比,U266细胞中CMTM5、caspase3和caspase9的mRNA表达水平与地西他滨呈浓度和时间依赖性,随浓度加大和时间延长表达水平增高更加显著(P值均<0.01);U266细胞中CMTM5表达水平与caspase3(r=0.937)、caspase9(r=0.945)呈正相关(P值均<0.001)。②过表达质粒转染的U266细胞,其CMTM5表达水平为133.10±10.35,较空白对照组(0.63±0.14)和质粒对照组(0.64±0.11)显著增高,差异有统计学意义(P值均<0.001);CMTM5过表达U266细胞组72 h细胞增殖活性为0.89±0.08,较对照组(1.32±0.02,t=5.005,P=0.008)和空载质粒组(1.30±0.03,t=4.700,P=0.009)显著降低,差异有统计学意义。结论MM细胞中CMTM5的缺失能够被去甲基化药物地西他滨逆转,其变化水平与caspase3、caspase9呈正相关。过表达CMTM5能够抑制U266细胞增殖。Objective To investigate the mechanism of chemokine-like factor superfamily member (CMTM) 5 on the proliferation of multiple myeloma cells.Methods RT-qPCR method was used to detect the expression and correlation of CMTM5,caspase3 and caspase9 in U266 after decitabine demethylation treatment;U266 transfected with pcDNA3.1 plasmid overexpressed CMTM5,then cell proliferation activity was detected by CCK-8 assay.Results Compared with the control group,the low-dose demethylation treatment increased mRNA expression of CMTM5,caspase3,and caspase9 in U266,and showed a time-dependent (P<0.01).The up-trend of CMTM5,caspase3,and caspase9 in the high-demethylation drug treatment group was more significant and also showed time-dependent (P<0.001);There was a significant positive correlation between CMTM5 and caspase3 (r=0.937) and caspase9 (r=0.945) in each group (P<0.001).After transfection of U266 with the pcDNA3.1-CMTM5 plasmid,overexpression of CMTM5 inhibited the cell proliferation activity compared with the control and pcDNA3.1-vector group.Conclusion Decitabine has a reductive effect on the low level of CMTM5 in U266 cells,and its recovery level is significantly positively correlated with caspase 3 and caspase9.Re-expression of CMTM5 inhibits the proliferative activity of U266.

关 键 词：多发性骨髓瘤 趋化素样因子超家族成员5 DNA甲基化 细胞凋亡 

分 类 号：R733.3[医药卫生—肿瘤]