COX2抑制剂联合KDM1A基因对肺癌A549细胞凋亡的诱导作用研究

Effects of COX2 Inhibitor Combined with KDM1A Gene on the Apoptosis of A549 Lung Cancer Cells in Vitro

目的探讨COX2抑制剂塞来昔布或KDM1A siRNA对肺癌A549细胞增殖凋亡的影响及其机制。方法以LipofectamineTM 2000为载体,将合成的KDM1A siRNA转染人肺腺癌A549细胞,Western blot检测转染后A549细胞中KDM1A的表达。塞来昔布或si-KDM1A处理A549细胞,分为NC组、塞来昔布组、si-KDM1A组、塞来昔布+si-KDM1A组。CCK8法检测细胞活力,流式细胞术检测细胞凋亡率;H2DCFHDA荧光探针检测ROS含量;Western blot检测STAT3、磷酸化STAT3及下游与增殖凋亡相关的基因PCNA和Bax蛋白表达。结果转染si-KDM1A后,A549细胞KDM1A表达明显受到抑制,与空白对照组比较,差异具有统计学意义(P<0.05)。塞来昔布和si-KDM1A均能明显抑制A549细胞活力,诱导细胞凋亡,提高细胞内ROS含量,下调p-STAT3和PCNA表达,上调Bax表达,与NC组比较,差异均具有统计学意义(P<0.05);且二者联合对细胞增殖、凋亡及STAT3信号通路的影响更明显,细胞活力、凋亡率、ROS含量及p-STAT3、PCNA、Bax表达水平与塞来昔布组和si-KDM1A组比较,差异均具有统计学意义(P<0.05)。结论 COX2抑制剂或KDM1A siRNA均可抑制肺癌A549细胞活力,诱导细胞凋亡,二者联用强于单独使用,其机制可能与提高细胞内ROS含量及抑制STAT3信号通路有关。

Objective To investigate the effect of COX2 inhibitor celecoxib or KDM1A siRNA on the proliferation and apoptosis of A549 lung cancer cells.Methods LipofectamineTM2000 was used as a carrier.KDM1A siRNA was transfected into human lung adenocarcinoma A549 cells,and the KDM1A expression in transfected A549 cells was detected by Western blot.After treated with celecoxib or si-KDM1A,the A549 cells were divided into NC group,celecoxib group,si-KDM1A group,celecoxib+si-KDM1A group.The cell viability was detected by CCK8 assay,and the apoptosis rate was detected by flow cytometry.The content of ROS was detected through the H2DCFHDA fluorescent probe.The expression of STAT3,p-STAT3,PCNA and Bax proteins were detected by Western blot.Results The expression of KDM1A in A549 cells after transfected with si-KDM1A was significantly inhibited,and there was significant difference when compared with the NC group(P<0.05).Celecoxib or si-KDM1A both could significantly inhibit A549 cell viability,induce apoptosis,increase the content of ROS in cells,down-regulate the expression of p-STAT3 and PCNA and up-regulate the expression of Bax.All differences were statistically significant when compared with NC group(P<0.05).The combination of celecoxib and si-KDM1A had much greater effects on cell proliferation,apoptosis and STAT3 signaling pathway.The results of cell viability,apoptosis rate,ROS content and expression of p-STAT3,PCNA and Bax were significantly different from those in celecoxib group and si-KDM1A group(P<0.05).Conclusion COX2 inhibitors or KDM1A siRNA both inhibit A549 cell viability and induce cell apoptosis.Their combination was stronger than single use.The mechanism may be related to the increase of intracellular ROS content and the inhibition of STAT3 signaling pathway.