miRNA-138-5p通过靶向调控CCNE1抑制黑色素瘤细胞增殖  

miRNA-138-5p inhibits melanoma cell proliferation by targeting CCNE1

作　　者：乔丽[1] 杨志勇[1] 李新技[2] 刘雯[1] QIA O Li YANG Zhiyong LI Xinji LIU Wen(Department of Dermatology, Air Force General Hospital, Beijing 100142, China)

机构地区：[1]空军总医院皮肤病医院,北京100142 [2]空军总医院肿瘤放疗科,北京100142

出　　处：《空军医学杂志》2017年第4期237-241,共5页Medical Journal of Air Force

摘　　要：目的研究miRNA-138-5p在黑色素瘤细胞增殖中的作用及其发挥作用的机制。方法 MTT法检测miRNA-138-5p转染后黑色素瘤细胞的增殖;Realtime PCR检测miRNA-138-5p转染后其靶基因CCNE1 mRNA水平的变化;Western blot检测miRNA-138-5p转染后CCNE1蛋白水平的变化;Luciferase Repor ter试验检测miRNA-138-5p对CCNE13'非翻译区(3'UTR)及其突变的影响;MTT法检测共转染miRNA-138-5p和CCNE1对黑色素瘤细胞增殖的影响。结果过表达miRNA-138-5p可抑制黑色素瘤细胞的增殖。CCNE1是miRNA-138-5p的一个功能性靶基因,转染miRNA-138-5p可以降低细胞中CCNE1的mRNA和蛋白水平。共转染miRNA-138-5p和CCNE1可以部分消除miRNA-138-5p对黑色素瘤细胞增殖的影响。结论 miRNA-138-5p通过其靶基因CCNE1在黑色素瘤细胞的增殖中发挥了重要的作用,miRNA-138-5p在黑色素瘤细胞中可能作为一个关键的药物靶点。Objective To study the role of miRNA-138-5p in the proliferation of melanoma cells and the mechanism. Methods MTT assay was used to detect the proliferation of melanoma cells after miRNA-138-5p transfection. Real-time PCR was used to detect the changes of the target gene CCNE1 mRNA after miRNA-138-5p transfection. Western blot was used to detect the changes of CCNE1 protein level after miRNA-138-5p transfection. Luciferase reporter assay was used to detect the effect of miRNA-138-5p on CCNE13＇s untranslated region （3＇UTR） and its mutations. MTT assay was used to detect the effect of co-transfection of miRNA-138-5p and CCNE1 on the proliferation of melanoma cells. Results Overexpression of miRNA-138-5p inhibited the proliferation of melanoma cells. CCNE1 was a functional target gene of miRNA-138-5p. Transfection of miRNA-138-5p could reduce the mRNA and protein levels of CCNE1 in cells. Co-transfection of miRNA-138-5p and CCNE1 could partially eliminate the effect of miRNA-138-5p on the proliferation of melanoma cells. Conclusion miRNA-138-5p plays an important role in the proliferation of melanoma cells through its target gene CCNE1, and miRNA-138-5p can be taken as a key drug target in melanoma cells.

关 键 词：MICRORNA 黑色素瘤 CCNE1 增殖 

分 类 号：R739.5[医药卫生—肿瘤]