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目的:研究去血清诱导人血管平滑肌细胞发生表型转化与microRNAs表达间的关系。方法:采取人血管平滑肌细胞克隆株HITASY,培养人血管平滑肌细胞(VSMCs)。用去血清的方法处理人血管平滑肌细胞,使VSMCs由去分化表型向分化表型转化,通过蛋白印迹法观察平滑肌细胞中分化标记物蛋白的变化,同时用实时定量PCR法检测细胞中相关microRNA的表达。结果:①去血清处理组与无去血清处理组比较,平滑肌细胞分化标记物(SM-α-actin、calponin)表达显著性增加,而通过SM-α-actin、calponin的蛋白表达量显著增加,提示血管平滑肌细胞向分化表型转化(P<0.05);②同时去血清处理组与无去血清处理组比较,Mir-649、Mir-944表达量显著增加,Mir-140、Mir-361表达量显著减少(P<0.05)。结论:Mir-649、Mir-944、Mir-140、Mir-361在血管平滑肌细胞表型转化中有关键性作用。
Objective: To study the relationship between phenotype change and microRNAs expression in serum-induced human vascular smooth muscle cells. Methods: Human vascular smooth muscle cell clone (HITASY) was used to culture human vascular smooth muscle cells (VSMCs). VSMCs were transformed from dedifferentiated phenotypes to differentiated phenotypes using human serum vascular smooth muscle cells. Western blotting was used to observe the changes of differentially expressed proteins in smooth muscle cells. Real-time quantitative PCR was used to detect the expression of microRNAs expression. Results: ① The expression of SM-α-actin (calponin) was significantly increased in serum-treated and untreated groups, while the expression of calponin was significantly increased by SM-α-actin, (P <0.05). ②The expression of Mir-649 and Mir-944 were significantly increased in the serum-treated group compared with the untreated group, the expression of Mir-140 and Mir-361 Significantly reduced (P <0.05). Conclusion: Mir-649, Mir-944, Mir-140 and Mir-361 are key factors in the phenotypic transition of vascular smooth muscle cells.