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目的探究维生素A对结肠癌细胞的增殖和凋亡的影响及相关的分子机制。方法 MTT检测不同浓度及不同处理时间的维生素A对结肠癌细胞SW620的影响,流式细胞计数仪检测维生素A对结肠癌细胞凋亡的影响,Western blotting检测维生素A处理后的SW620细胞中AKT/FOXO3a信号通路的变化及促凋亡蛋白Bim的表达,FOXO3a特异性沉默后检测细胞凋亡的变化及Bim的表达。结果维生素A处理后的SW620细胞数量明显减少,且具有时间和浓度依赖性。流式细胞计数结果进一步验证了维生素A的促凋亡效应。Western blotting结果显示维生素A能抑制AKT和OXO3a的磷酸化,导致FOXO3a在核内聚积,促进Bim的表达。当siRNA干扰SW620细胞中FOXO3a的表达,发现维生素A的促凋亡作用在FOXO3a沉默后的SW620细胞中不明显,Bim的表达也显著下调。结论 Bim作为FOXO3a的下游分子,AKT-FOXO3a信号通路对Bim的调控作用介导了维生素A的促凋亡效应。
Objective To investigate the effects of vitamin A on the proliferation and apoptosis of colon cancer cells and the related molecular mechanisms. Methods MTT was used to detect the effect of vitamin A on colon cancer cell line SW620 at different concentrations and different treatment time. Flow cytometry was used to detect the effect of vitamin A on the apoptosis of colon cancer cells. Western blotting was used to detect the AKT / The changes of FOXO3a signal pathway and the expression of Bim, the apoptosis-inducing protein and the expression of Bim were detected by FOXO3a-specific silencing. Results The number of SW620 cells treated with vitamin A significantly decreased, with a time-and concentration-dependent manner. Flow cytometry results further verify the pro-apoptotic effect of vitamin A. Western blotting showed that vitamin A can inhibit the phosphorylation of AKT and OXO3a, leading to the accumulation of FOXO3a in the nucleus and promoting the expression of Bim. When siRNA interfered with FOXO3a expression in SW620 cells, it was found that the pro-apoptotic effect of vitamin A was not obvious in SW620 cells after FOXO3a silencing, and the expression of Bim was also significantly down-regulated. Conclusions Bim, a downstream molecule of FOXO3a, mediates the pro-apoptotic effect of vitamin A through the regulation of Bim by AKT-FOXO3a signaling pathway.