microRNAs在恶性黑色素瘤B16F0和B16F10细胞中的差异表达  

Identification of microRNAs differentially expressed in malignant melanoma B16F0 and B16F10 cells by microRNAs microarray

作　　者：张彬彬[1] 张亚平[1] 丁琼琼[1] 杨海杰[1] 马建[1] 冯志伟[1] 王磊[1] 

机构地区：[1]新乡医学院生命科学技术学院,河南新乡453003

出　　处：《南京医科大学学报（自然科学版）》2017年第7期823-829,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：NSFC-河南联合基金(U1604166);新乡医学院研究生科研创新支持计划(YJSCX201517Y)

摘　　要：目的:探讨及分析黑色素瘤细胞B16F0和B16F10中microRNAs(miRNAs)表达差异性,筛选与恶性黑色素瘤增殖、迁移和侵袭相关的miRNAs。方法:通过体内和体外实验分别验证B16F0和B16F10细胞的差异,并通过Ribo Array TMmi DETECTTMMouse Array分析小鼠黑色素瘤B16F0和B16F10细胞中miRNAs表达的差异性,筛选出与恶性黑色素瘤相关的miRNAs,并应用real-time PCR验证筛选结果的可信性。结果:B16F10细胞比B16F0具有更强的迁移和增殖能力,芯片结果显示,与黑色素瘤B16F0细胞相比,在B16F10细胞中有38个miRNAs存在表达差异,其中15个miRNAs表达上调,23个miRNAs表达下调,差异均具有统计学意义。Real-time PCR对部分miRNAs进行验证,结果与芯片分析一致,其中miR-26a-1-3p、miR-28b、miR-29b-3p、miR-24-2-5p表达显著上调,而miR-763、miR-431-5p、miR-205-5p、let-7c-2-3p表达显著下调。结论:差异表达的miRNAs与恶性黑色素瘤的发生、发展可能存在密切相关,可能为恶性黑色素瘤的早期诊断及治疗提供有力的靶向依据。Objective：To analyze the differential expression profile of microRNAs in murine melanoma B16F0 and B16F10 cells, and to isolate the microRNAs related to the proliferation and metastasis of malignant melanoma. Methods：We verified the difference of B16F10 and B16F0 cell in vivo and in vitro. And then RiboArrayTM miDETECTTMMouse Array was adopted to compare the mi- croRNAs expressed in murine melanoma B16FO and B16F10 cells. The differently expressed microRNAs were further confirmed by real time PCR. Results：B16FlO cells had stronger abilities of migration and proliferation in vivo and in vitro. Further more, compared with B16FO eells,a total number of 38 microRNAs were statically differently expressed in B16F10 cells,with 15 upregulated and 23 downregulated. Consistent with RiboArray results,the expressions of miR-26a-l-3p,miR-28b,miR-29b-3p, and miR-24-2-Sp were sig- nificantly upregulated and the expressions of miR-763,miR-431-Sp,miR-205-Sp,and let-7c-2-3p were markedly downregulated re- vealed by real time PCR. Conclusion： Our findings suggest that the differentially expressed microRNAs were related to the occurrence and development of malignant melanoma, and might serve as novel targets for early diagnosis and treatment of melanoma.

关 键 词：恶性黑色素瘤 MICRORNAS 表达谱 REAL-TIME PCR 

分 类 号：Q28[生物学—细胞生物学]