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目的探讨miR-33a对人皮肤恶性黑色素瘤系A375生物学功能的影响,分析其可能机制。方法脂质体转染人工合成microRNA片段mimics,干预miR-33a在人皮肤恶性黑色素瘤细胞系A375细胞中的表达水平,行实时PCR检测干预效能。行MTT法检测细胞增殖能力;行细胞黏附实验检测其黏附能力;行细胞划痕实验、Transwell实验检测其迁移侵袭能力;以表达谱基因芯片筛选miR-33a过表达后细胞差异表达基因。结果检测发现,miR-33a过表达组细胞增殖、黏附、迁移侵袭能力均明显弱于空白对照组及阴性对照组,差异有统计学意义(P<0.05);miR-33a过表达后210个基因表达出现变化,包括84个上调、126个下降,细胞信息学分析指出差异表达基因广泛参与细胞周期调控、转录调控。结论 miR-33a对人皮肤恶性黑色素瘤细胞系A375增殖、黏附、迁移能力均有一定抑制作用,这与其导致的基因表达谱改变有关。
Objective To investigate the effect of miR-33a on the biological function of human skin malignant melanoma line A375 and to analyze its possible mechanism. Methods Liposomes were transfected into mimics, a synthetic microRNA fragment, to interfere the expression of miR-33a in human melanoma cell line A375. Real-time PCR was used to detect the efficacy of miR-33a. Cell adhesion assay was performed by MTT assay. Cell adhesion assay was performed by cell adhesion assay. Scratch assay and Transwell assay were used to detect the migration and invasion ability. The differentially expressed genes were screened by miR-33a over expression. The results showed that miR-33a over-expression cells proliferation, adhesion, migration and invasion were significantly weaker than the control group and the negative control group, the difference was statistically significant (P <0.05); miR-33a over-expression of 210 genes Changes in expression appeared, including 84 up-regulated and 126 down-regulated. Cellular informatics analysis indicated that differentially expressed genes were widely involved in cell cycle regulation and transcriptional regulation. Conclusion miR-33a inhibits the proliferation, adhesion and migration of human dermal malignant melanoma cell line A375, which is related to the change of gene expression profile induced by miR-33a.