FDCs-miR-548m-CDK6轴在套细胞淋巴瘤集落形成中的研究  

Role of FDCs-miR-548m-CDK6 axis in clonogenicity of mantle cell lymphoma

作　　者：王芳[1] 张新伟[1] 张翼鷟[2] 魏枫[1] 任秀宝[1] 

机构地区：[1]天津医科大学肿瘤医院肿瘤研究所免疫研究室,国家肿瘤临床医学研究中心,天津市肿瘤免疫与生物治疗重点实验室,天津市300060 [2]天津医科大学肿瘤医院血液科,天津市300060

出　　处：《中国肿瘤临床》2014年第18期1175-1179,共5页Chinese Journal of Clinical Oncology

基　　金：国家重点基础研究发展计划(973计划)项目(编号:2012CB9333004);天津市自然科学基金项目(编号:14JCYBJC27100)资助~~

摘　　要：目的:探讨FDCs-miR-548m-CDK6轴在套细胞淋巴瘤(mantle cell lymphoma,MCL)集落形成中的作用。方法:分别采用RT-qPCR和Western blot检测MCL细胞与滤泡树突状细胞(FDCs)共培养后miR-548m和CDK6的变化。以生物信息学软件预测miR-548m的靶点,Western Blot检测MCL细胞系分别转染pre-miR-548m和anti-miR-548后细胞周期蛋白依赖激酶6(CDK6)的变化。荧光素酶报告基因实验验证CDK6是否为miR-548m的直接作用靶点。MCL细胞系与/不与FDCs共培养,过表达miR-548m或者敲低CDK6后MCL的集落形成能力。结果:FDCs与MCL的粘附作用可以下调miR-548m并上调CDK6。生物信息学软件预测显示CDK6的3'UTR是miR-548m的潜在靶点,且过表达miR-548m能够减少CDK6的表达,抑制miR-548m表达能够增加CDK6。荧光素酶报告基因实验证实CDK6的3'UTR是miR-548m的一个直接作用靶点。集落形成实验显示过表达miR-548m或者敲低CDK6后,细胞的集落形成能力明显减弱。结论:FDCs通过抑制MCL中miR-548m表达上调CDK6,增强MCL的集落形成能力。Objective：To study the role of FDCs-miR-548m-CDK6 axis on clonogenicity in mantle cell lymphoma. Methods：RT-qPCR and Western blot were used respectively to test the expression of miR-548m and CDK6. Bioinformatics assay was applied to predict the targets of miR-548m, and Western Blot was used to test the expression level of CDK6 after miR-548m overexpression or in-hibition. Luciferase report assay was performed to test whether CDK6 was a direct target of miR-548m. Colony forming assay was used to test the colony forming activity in MCL after overexpression of miR-548m or knockdown of CDK6. Results：Cell adhesion to FDCs induced downregulation of miR-548m and CDK6 expression in MCL. Bioinformatics assay revealed that miR-548m could target the 3＇-UTR of CDK6 and that a negative correlation exists between the level of miR-548m and the CDK6 expression. Luciferase report as-say confirmed that miR-548m directly targeted 3＇-UTR of CDK6. Colony forming assay showed that overexpression of miR-548m or knockdown of CDK6 significantly suppressed MCL colony formation. Conclusion：This study reveals that FDC-enhanced mantle cell lymphoma clonogenicity is mediated by the miR-548m/CDK6 axis.

关 键 词：套细胞淋巴瘤 滤泡树突状细胞 miR-548m 细胞周期蛋白依赖性激酶6 集落形成 

分 类 号：R733.1[医药卫生—肿瘤]