氯胺酮对人肺癌A549细胞生长的影响及相关机制研究

The effect and mechanism of ketamine on proliferation of human lung adenocarcinoma A549 cells

目的 :探讨麻醉类药物氯胺酮和吗啡对人肺癌A549细胞生长的影响及相关作用机制。方法:体外培养肺癌细胞系A549,细胞培养24 h后,随机分为3组:对照组(NC组,加0.9%Na Cl处理)、氯胺酮组(K1、K2、K3组,分别加氯胺酮100、200、400μg/m L处理)和吗啡组(M1、M2、M3组,分别加吗啡25、50、100μg/m L处理)。持续作用24 h后,Transwell小室培养体系测定肿瘤细胞的迁移率;双荧光素酶报告基因法测定NF-κB转录活性;48 h后MTT法测定细胞增殖抑制率;Western blot技术测定NF-κB P65的蛋白表达;QPCR技术检测细胞内IL-1β和COX-2的m RNA表达。结果:1与NC组比较,M1、M2组表现为促进A549细胞迁移(P<0.05),M3组作用不明显;K1、K2、K3组表现为抑制A549细胞迁移(P<0.05);2与NC组比较,M1、M2组表现为促进A549细胞增殖(P<0.05),M3组作用不明显;K2、K3组表现为抑制A549细胞增殖(P<0.05),K1组不明显;3与NC组比较,M1、M2、M3组表现为促进IL-1β、COX-2的m RNA表达(P<0.05);K2、K3组表现为抑制IL-1β、COX-2的m RNA表达(P<0.05),K1组不明显;4与NC组比较,M1、M2、M3组表现为促进P65的转录和蛋白表达(P<0.05),K1、K2、K3组表现为抑制P65的转录和蛋白表达(P<0.05)。结论:与阿片类药物不同,氯胺酮通过NF-κB信号通路及其下游因子IL-1β和COX-2抑制人肺腺癌细胞A549的迁移和增殖,可能更适用于肺癌患者的术后镇痛。

Objective：To investigate the effect and mechanism of anesthetic ketamine and morphine on the proliferation of cultured human lung adenocarcinoma A549 cell. Methods：Human lung adenocarcinoma A549 cells were treated with different concentration of ketamine（K1,K2,K3 is 100,200,and 400 μg / m L,respectively）and morphine（M1,M2,M3 is 25,50 and 100 μg / m L,respectively）for24 and 48 h. After cultured for 24 h,the effect of ketamine and morphine on the invasive ability of A549 cells were measured by transwell invasion. NF-κB transcription activity were determined by luciferase assay. After cultured for 48 h,the proliferation inhibition rates of A549 cells were assessed by methyl thiazolyl tetrazolium（MTT） assay. Protein expressions of NF-κB P65 were determined by Western blot. IL-1β and COX-2 expression were determined by QPCR. Results：（1）Compared with the control group,M1 and M2 enhanced A549 cell proliferation and migration（P 〈0.05）,while K1,K2 and K3 suppressed A549 cells migration（P 〈0.05）.（2）M1,M2 and M3 enhanced IL-1β and COX-2 expression（P〈 0.05）,but K2 and K3 had opposing effects（P 〈0.05）. M1,M2,and M3 enhanced NF-κB and P65 transcription activity and protein expressions（P 〈0.05）,while K1,K2 and K3 had opposing effects（P 〈0.05）. Conclusion：Unlike opioids,ketamine inhibits the proliferation and migration of A549 through NF-κB and its down stream factor IL-1β and COX-2,may be more suitable for lung cancer patients postoperative analgesia.