摘要
目的:探讨黑素瘤中microRNA(miRNA)-183/96/182同源簇的表达丰度及可能的调控机制。方法:采用荧光定量聚合酶链反应(qPCR)检测黑素瘤细胞系(A375、WM35)与黑素瘤样本中miR-183/96/182的表达丰度与差异。JASPAR数据库分析miR-183/96/182启动子区域可能存在的转录因子结合位点。采用荧光素酶报告基因实验与染色质免疫沉淀法(ChIP)分析转录因子对miR-183/96/182表达的调控机制。分析转录因子与miR-183/96/182对黑素瘤细胞迁移的影响。结果:miR-183/96/182同源簇中,miR-182在黑素瘤细胞系与肿瘤样本中的表达较其他两种miR升高(P<0.05)。黑素瘤细胞系过表达TEAD1后,miR-182的表达升高(P<0.05);抑制miR-182后,TEAD1诱导的黑素瘤细胞转移的作用降低(P<0.01)。结论:miR-182在黑素瘤中高表达,其可能通过TEAD1的转录调控参与黑素瘤细胞转移。
Objective:To explore the expression level and regulation mechanisms of microRNA (miR)-183/96/182 cluster in melanoma cells. Methods:qPCR was used to detect the expression and difference of miR-183/96/182 in melanoma cell lines (A375, WM35) and melanoma clinical samples. JASPAR database was analyzed to investigate the potential transcription factor binding sites in the promoter region of miR-183/96/182. Luciferase assay and chromatin immunoprecipitation assay were used to explore the regulatory mechanism of transcription factors on miR-183/96/182 expression. Cell migration assay was performed to evaluate the effects of transcription factors and miR-183/96/182 on the metastasis of melanoma cells. Results:The expression of miR-182 of the miR-183/96/182 homologous cluster was higher in melanoma cell lines and tumor samples compared with miR-183 and miR-96 ( P <0.05). The expression of miR-182 increased with TEAD1 overexpressing in melanoma cell lines ( P<0.05). The metastasis of melanoma induced by TEAD1 reduced with miR-182 expression inhibition ( P<0.01). Conclusions:miR-182 is highly expressed in melanoma cells, and it might play an important role in the metastasis of melanoma via the transcriptional regulation of TEAD1.
作者
陈诚
张学军
CHEN Cheng;ZHANG Xue-jun(Department of Plastic Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, China)
出处
《中国临床医学》
2019年第1期89-92,共4页
Chinese Journal of Clinical Medicine
