白藜芦醇与吉非替尼的协同抗肺癌作用及机制研究

Synergistic Effects between Resveratrol and Gefitinib on Lung Cancer

目的探讨白藜芦醇是否能提高吉非替尼对肺癌细胞的杀伤活性并研究其机制。方法MTT法检测人非小细胞肺癌细胞系PC9细胞在低浓度吉非替尼和白藜芦醇处理下的细胞活力。Western blot实验检测低浓度吉非替尼和白藜芦醇对PC9细胞c-met表达水平,表皮生长因子受体(EGFR)、磷脂酰肌醇激酶(PI3K)、丝氨酸苏氨酸激酶(AKT)磷酸化水平及caspases活化水平。流式细胞术检测PC9细胞在低浓度吉非替尼和白藜芦醇处理下的凋亡率。结果低浓度吉非替尼联合白藜芦醇对PC9的细胞活力抑制率显著高于低浓度吉非替尼组和白藜芦醇组[(57.7±3.3)%比(12.4±1.1)%、(8.6±0.8)%,P<0.05]。白藜芦醇处理可诱导PC9细胞c-met蛋白的下调并显著增强低浓度吉非替尼对PC9细胞PI3K和AKT磷酸化水平的抑制作用。低浓度吉非替尼联合白藜芦醇组对PC9细胞活力的抑制率和凋亡诱导率显著高于低浓度吉非替尼组和低浓度吉非替尼+白藜芦醇+c-met质粒组[抑制率:(57.7±3.3)%比(12.4±1.1)%、(16.3±1.3)%,P<0.05;凋亡率:(30.9±1.8)%比(7.2±0.5)%、(10.6±0.8)%,P<0.05]。低浓度吉非替尼联合白藜芦醇组对PC9细胞caspase-9及caspase-3的活化显著强于低浓度吉非替尼组和低浓度吉非替尼+白藜芦醇+c-met质粒组。结论白藜芦醇可能通过下调c-met的表达提高肺癌细胞对吉非替尼的敏感性。

Objective To investigate the role of resveratrol in increasing the sensitivity of lung cancer cells to gefitinib and the underlying mechanisms. Methods MTT assay was performed to evaluate the viability of PC9 cells treated with resveratrol and low dose of gefitinib. Western blot was used to detect the expression of c-met, phosphorylation of EGFR, PI3K and AKT and activation of caspases on PC9 ceils treated with resveratrol and low dose of gefitinib. Flow cytometry analysis was carried out to measure the apoptotic rate of PC9 cells treated with resveratrol and low-dose of gefitinib. Results Cell viability inhibitory rate of PC9 cells in low-dose gefitinib plus resveratrol was significantly higher than the low-dose gefitinib group and resveratrol group（57.7%±3.3% vs 12.4%±1.1%, 8.6%±0.8%, P〈0.05）. Treatment with resveratrol significantly downregulated the expression of c-met as well as enhancing the inhibition of PI3K and AKT phosphorylation induced by gefitinib. Cell viability inhibitory rate and apoptotic rate of PC9 cells in low-dose gefitinib plus resveratrol group was significantly higher than that in the low-dose gefitinib group and the low-dose gefitinib＋resveratrol＋c-met plasmid group（inhibitory rate： 57.7%±3.3% vs 12.4%±1.1%, 16.3%±1.3%; apoptotic rate： 30.9%±1.8% vs 7.2%±0.5%, 10.6%±0.8%; P〈0.05）. Activation of cas- pase-9 and caspase-3 in the low-dose gefitinib plus resveratrol group was more remarkable than that in the low- dose gefitinib group and the low-dose gefitinib＋resveratrol＋c-met plasmid group in PC9. Conclusion Resveratrol can increase the sensitivity of lung cancer cells to gefitinib by downregulating the expression of c-met.