下调microRNA-192基因表达对恶性黑素瘤A375细胞增殖及早期凋亡的影响

The effect of knockdown of microRNA-192 gene on the proliferation and apoptosis in the A375 melanoma cell line

目的:研究microRNA-192基因表达降低与恶性黑素瘤A375细胞增殖及凋亡的关系。方法:除空白对照组外,运用脂质体介导法将空载体和为microRNA-192设计的small interfering RNA(si RNA)转染到恶性黑素瘤A375细胞中,并利用实时定量PCR检测microRNA-192的表达。通过形态学观察、细胞计数法、3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法以及流式细胞免疫学等方法,分析空白对照组、空载体对照组(lipofectamine2000)和实验组恶性黑素瘤A375细胞的增殖和早期凋亡情况。结果:空白对照组和空载体对照组之间相比较,细胞增殖和凋亡情况几乎一致,差异无统计学意义。与空载体对照组相比,实验组存活的恶性黑素瘤A375细胞的活性和数量明显降低,此现象并随着si RNA浓度的提高而愈发明显。另外,实验组恶性黑素瘤A375细胞细胞凋亡指数较空载体对照组明显升高。结论:通过下调恶性黑素瘤细胞中microRNA-192基因表达可能会有效地抑制恶性黑素瘤的生长并促使其细胞凋亡。

Objective： We assessed whether microRNA-192 deficiency influences the proliferation and apoptosis in A375 melanoma cell line. Methods： A375 melanoma cells were transfected with lipofeetamine alone or small interfering RNA（MicroRNA-1192 siR NA）-loaded lipofectamine. A group of untreated cells served as an additional normal control. The expression of mieroRNA-192 was analyzed with real-time PCR Cell proliferation and early apoptosis were assessed with inverted microscope, MTI＇ assay, and flow cytometry. Result： There were no significant differences in proliferation and apoptosis between normal controls and cells-treated with lipofectamine alone. In comparison with ]ipofectamine alone, siRNA treatment induced significant reduction in cell activity and the number of the A375 melanoma cells in a dose-dependent manner. Additionally, transfection with MicroRNA-1192 siRNA dramatical- ly increased apoptosis index. Conclusion： Downregulation of microRNA-192 can effectively inhibit the proliferation and accelerate the apoptosis in A375 melanoma cell line.