辛伐他汀通过JAK2/STAT3通路诱导肺癌A549细胞凋亡的研究

Simvastatin induced apoptosis in A549 lung cancer cell by JAK2/STAT3 pathway

目的探究辛伐他汀对非小细胞肺癌A549细胞凋亡的影响及作用的可能机制。方法用辛伐他汀处理人肺癌A549细胞后,采用CCK8法检测辛伐他汀对A549细胞活性的影响,通过流式细胞术检测细胞周期,TUNEL法检测对A549细胞凋亡的影响,Western Blot法检测细胞凋亡相关蛋白Bcl-2、Bax的表达,EMSA法检测辛伐他汀对STAT3/DNA结合活性的影响,Western Blot法检测JAK2/STAT3相关蛋白的表达。结果与对照组细胞相比,辛伐他汀处理组的A549细胞的活性受到抑制(P<0.05),并且随着药物处理浓度的升高和处理时间的延长逐渐降低;采用IC50浓度(40μg/ml)辛伐他汀处理A549细胞48 h,可引起A549细胞周期阻滞在G0/G1期,与对照组比较,差异有统计学意义(P<0.01),TUNEL法结果显示辛伐他汀能引起A549细胞的凋亡;辛伐他汀处理组凋亡蛋白Bax表达量升高,凋亡抑制蛋白Bcl-2表达降低(P<0.05);辛伐他汀可使STAT3/DNA结合活性降低,p-JAK2、p-STAT3蛋白表达降低(P<0.01)。结论辛伐他汀可能通过抑制JAK2/STAT3信号通路诱导肺癌A549细胞的凋亡。

Objective To investigate the effects of simvastatin on apoptosis in A549 non-small cell lung cancer (NSCLC) cell and its possible mechanism.Methods Human lung cancer A549 cells were treated with simvastatin, and CCK8 method was used to detect the influence of simvastatin on A549 cell activity. The cell cycle was detected by flow cytometry, and apoptosis in A549 cells was detected by terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) assay. The expression of cell apoptosis related proteins, Bcl-2 and Bax were detected by Western Blot method. Electrophoretic mobility shift assay method was used for detecting the simvastatin influence on STAT3/DNA binding activity, and JAK2/STAT3 protein expression were detected by Western Blot.Results Compared with the control group, the activity of A549 cells in the simvastatin treatment group was inhibited (P<0.05), and gradually decreased with the increase of drug treatment concentration and the extension of treatment time. A549 cells were treated with simvastatin at IC50 concentration (40 g/ml) for 48 h, causing A549 cell cycle arrest in G0/G1 phase, and the difference was statistically significant, compared with the control group (P<0.01). TUNEL assay showed that simvastatin could induce apoptosis in A549 cells.The expression level of apoptotic protein Bax, increased, and the expression level of apoptotic inhibitory protein bcl-2 decreased in the simvastatin treatment group (P<0.05). Simvastatin reduced the binding activity of STAT3/DNA and the expression of p-JAK2 and p-STAT3 (P<0.01).Conclusions Simvastatin may induce apoptosis of A549 lung cancer cells by inhibiting the JAK2 /STAT3 signaling pathway.