硼替佐米对A549细胞增殖及p21、p27表达的影响  

Anti-proliferation effect of proteasome inhibitor bortezomib in human lung adenocarcinoma A549 cells

作　　者：谢燕[1] 喻秀丽[1] 童立纺[1] 

机构地区：[1]重庆医科大学附属第一医院青杠老年护养中心,重庆400016

出　　处：《第三军医大学学报》2012年第17期1775-1778,共4页Journal of Third Military Medical University

摘　　要：目的研究蛋白酶体抑制剂硼替佐米对人肺癌A549细胞增殖、细胞周期及在体成瘤的影响。方法采用不同浓度的硼替佐米(0、100、200 nmol/L)作用于A549细胞4 h,观察其对蛋白酶体活性的影响。用硼替佐米(0、100、200 nmol/L)处理A549细胞48 h,采用MTT法、3H-TdR掺入法检测细胞增殖;流式细胞术PI染色检测细胞周期;Westernblot检测p21、p27表达水平的变化;连续注射观察硼替佐米对裸鼠A549细胞移植瘤体积及质量的影响。结果硼替佐米可显著抑制A549细胞的蛋白酶体活性。MTT法、3H-TdR法检测显示,100 nmol/L处理组细胞较对照组增殖率分别降低24.5%、29.5%,200 nmol/L处理组进一步显著下降(P<0.05);细胞周期分析显示,100、200 nmol/L处理组G0/G1期较对照组明显增加(P<0.05)。Western blot检测显示,48 h时100、200 nmol/L处理组p21、p27蛋白表达较对照组明显增加,且呈浓度依赖(P<0.05),48 h蛋白质水平高于24 h。连续注射硼替佐米后裸鼠移植瘤的质量抑制率达(36.0±12.4)%。结论硼替佐米可抑制A549细胞增殖,诱导细胞凋亡及发生G0/G1期阻滞,在体注射可抑制裸鼠成瘤,其抗肿瘤活性可能与抑制细胞周期蛋白p21、p27的降解有关。Objective To investigate the effects of a proteasome inhibitor bortezomib on the proliferation,cell cycle and tumor formation of human lung adenocarcinoma A549 cells,and to explore the possible mechanisms involved.Methods Human lung adenocarcinoma A549 cells were cultured and treated with different concentrations of bortezomib（0,100 and 200 nmol/L） for 4 h,and the proteasome activity were measured to confirm its inhibitory effects.The A549 cells were further treated with different concentrations of bortezomib（0,100 and 200 nmol/L） for 24-48 h.Cell proliferation was determined by MTT assay and 3H-TdR absorption,respectively.Cell cycle was detected by flow cytometry,and the protein levels of p21 and p27 were determined by Western blotting.A549 cells were implanted subcutaneously into nude mice to establish the xenograft tumor models,and the xenograft tumor bearing mice were subjected to bortezomib.The tumor volume and weight in all groups were measured,and the tumor inhibitory rate was calculated.Results Bortezomib significantly inhibited the proteasome activity in A549 cells.The MTT and 3H-TdR results showed that cell proliferation decreased by 24.5% and 29.5% in the 100 nmol/L bortezomib group,respectively,and a more significant decease was observed in the 200 nmol/L bortezomib group compared with the control group（P0.05）.Cell cycle analysis showed that the number of cells at G0/G1 phase was significantly increased in the 100 nmol/L and 200 nmol/L bortezomib groups compared with the control group（P0.05）.Western blot results showed that p21 and p27 significantly increased in a dose-dependent manner in the 100 nmol/L and 200 nmol/L bortezomib groups compared with the control group（P0.05）.In addition,the levels of p21 and p27 in the bortezomib group were significantly higher at 48 h than those at 24 h.After continuous injection,the weight of the xenograft tumors was reduced by（36.0±12.4）% in the bortezomib group compared with the control group.Conclusion Bortezomib can inhibit A549 cell prol

关 键 词：硼替佐米 肺癌 细胞周期 

分 类 号：R73-361[医药卫生—肿瘤] R730.23[医药卫生—临床医学]