二甲双胍对肺癌A549细胞增殖、迁移、侵袭能力及乳酸生成的影响

The Effect of Metformin on Proliferation,Migration,Invasion and Lactic Acid Secretion of Lung Cancer A549 Cells

[目的]探讨二甲双胍对肺癌A549细胞增殖、迁移、侵袭能力、乳酸生成的影响。[方法]应用Annexin V/PI和API等标记,流式细胞仪对肺癌A549细胞凋亡及周期特异性的检测,运用RT-PCR的方法检测Caspase-3、Caspase-9活化情况。MTT检测细胞增殖活力;Transwell体外迁移实验检测细胞体外迁移和侵袭能力;并检测乳酸生成情况。[结果 ]二甲双胍诱导的细胞凋亡主要在细胞周期的G1期发生。采用二甲双胍凋亡诱导,对照组未做处理,结果显示观察组的Caspase-3 m RNA（1.12±0.56）、Caspase-9 m RNA（1.10±0.29）的表达值显著高于对照组（0.82±0.31,0.72±0.26）。Transwell结果显示,第7d二甲双胍组体外迁移细胞数（2.69±0.56）明显增加;二甲双胍组细胞活力（52.39%±5.96%）下降;二甲双胍组乳酸生成（12.89±1.03）明显降低。[结论 ]二甲双胍诱导的细胞凋亡与肺癌A549细胞的细胞周期相关,存在G1期细胞周期阻滞的周期特异性,且此过程中发生了caspase活化,抑制细胞活性及迁移侵袭能力,降低乳酸的生成,促进肿瘤细胞凋亡。

[Objective] To observe the effect of metformin on proliferation,migration,invasion and lactic acid secretion of lung cancer A549 cells. [Methods] Apoptosis and specificity of lung cancer A549 cells were detected by Annexin V/PI and API. The activation of Caspase-3 and Caspase-9 by RT-PCR was detected by Flow cytometry. The cell viability was detected by MTT assay. The migration and invasion of cells in vitro was detected by Transwell migration assay. And the lactate secretion was detected. [Results] Metformin-induced apoptosis occurs mainly in the G1 phase of the cell cycle. The expression of Caspase-3（1.12±0.56） and Caspase-9（1.10±0.29） m RNA in the observation group was significantly higher than those in the control group（0.82±0.31,0.72±0.26）.Transwell results showed that the number of cells migrating in the metformin group（2.69±0.56） was significantly increased on day 7;the cell viability 52.39%±5.96% decreased in the metformin group and the lactic acid production（12.89 ±1.03） in the metformin group was significantly decreased.[Conclusion] Metformin-induced apoptosis is related to the cell cycle of lung cancer A549 cells.There is a period-specificity of cell cycle arrest in G1 phase,and Caspase activation occurs during the process,which inhibits cell activity and migration ability,reduces lactate secretion,involved in apoptosis.