鸡血藤黄酮类有效部位SSCE对人肺腺癌细胞A549细胞周期调控的分子机制

Molecular Mechanism of Cell Cycle Regulation of Spatholobus Suberectus Flavonoids Effective Part (SSCE) on the A549 Human Lung Adenocarcinoma Cell

目的:探讨鸡血藤黄酮类有效部位(SSCE)对人肺癌A549细胞的周期阻滞作用及其分子机制。方法:采用CCK-8法检测SSCE对A549细胞的增殖影响;流式细胞仪检测细胞周期;RT-PCR法检测CDK1、2、4、6mRNA表达;Western-blot法测定CDK1、2、4、6蛋白表达。结果:SSCE抑制A549细胞的增殖,SSCE引起A549细胞G1/S期和/或G2/M期阻滞。160mg/L SSCE作用A549细胞24h后,CDK1、CDK2、CDK6mRNA表达较对照组减少(P<0.05);CDK1、CDK2、CDK6蛋白含量较对照组低(P<0.05);160mg/L SSCE作用A549细胞48h后,细胞内CDK1mRNA表达较对照组减少(P<0.05);细胞内CDK1蛋白含量较对照组低(P<0.05)。结论:SSCE通过下调CDK1mRNA转录水平及蛋白表达量引起A549细胞G2/M期阻滞;通过下调CDK2、6mRNA转录水平及蛋白表达量引起A549细胞G1期阻滞。

Objective：To study the effects of spatholobus suberectus flavonoids（SSCE） on cell cycle arrest and its molecular mechanism.Methods：The CCK-8 assay was used to detect the A549 proliferation;flow cytomytry was adopted to observe the cell cycle arrest;the expression of CDK1、2、4、6、mRNA was tested by RT-PCR method,expression of CDK1、2、4、6 protein was determined by Western blotting method.Results：SSCE can obviously reduce the A549 cells quantity.SSCE acting on A549 cells caused G1 /S phase and/or G2 /M phase arrest.Expression of CDK1、CDK2、CDK6 mRNA in 160mg/L SSCE- 24h-treated group was significantly higher than that of control group（P 0.05）;content of CDK1、CDK2、 CDK6 protein in 160mg/L SSCE-24h-treated group was increased compared with control group （ P 0.05）,Expression of CDK1 mRNA in 160mg/L SSCE-48h-treated group was significantly higher than that of control group （ P 0.05）;content of CDK1 protein in 160mg/L SSCE-48h-treated group was increased compared with control group（P 0.05）.〗Conclusion：SSCE by down-regulating CDK1 mRNA transcription and protein expression in A549 cells caused G2 /M phase arrest;SSCE by down-regulating CDK2,6 mRNA transcription and protein expression in A549 cells,causing G1 phase arrest.