蛋白酶体抑制剂硼替佐米对A549细胞的抗肿瘤.docVIP

硼替佐米对A549细胞增殖及p21、p27表达的影响 谢燕，喻秀丽，童立纺（400016 重庆，重庆医科大学附属第一医院青杠老年护养中心） [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，并进一步探讨其作用机制。方法 采用不同浓度的硼替佐米（0、100nmol/L、200nmol/L）作用于A549细胞4h，观察其对蛋白酶体活性的影响。进而用硼替佐米（0、100nmol/L、200nmol/L）处理A549细胞48h，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；Western blot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细胞移植瘤体积及重量的变化。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实验显示，100nmol/L处理组细胞较对照组增殖率分别降低24.5%、29.5%，而200nmol/L处理组则进一步显著下降（P0.05）；细胞周期分析显示， 100nmol/L、200nmol/L处理组G0/G1期比例较对照组明显增加（P0.05）。Western blot结果发现，48h时100nmol/L、200nmol/L处理组p21、p27蛋白表达较对照组明显增加，且呈浓度依赖（P0.05），且48h组蛋白质水平高于24h时。连续注射硼替佐米后裸鼠移植瘤的重量抑制率达36%。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。 [关键词] 硼替佐米；肺癌；细胞周期 Anti-proliferation Effects of the Proteasome Inhibitor Bortezomib in Human Lung Adenocarcinoma A549 Cells Xie Yan，Yu Xiuli，Tong Lifang (Department of Qinggang Geriatrics，Frist Affiliated Hospital of Chongqing Medical University，400016，Chongqing，China) [Abstract] Objective To investigate the anti-tumor effect of bortezomib, a proteasome inhibitor, in human lung adenocarcinoma A549 cells and to explore the possible mechanism involved. Methods Human lung adenocarcinoma A549 cells were cultured and then treated with bortezomib (0, 100, 200nmol/L) for 4h, the proteasome activity were measured to confirm its inhibitory effects. A549 cells were further treated with different concentrations of bortezomib (0, 100, 200nmol/L) for 24-48h. Cell proliferation was determined by MTT assay and 3H-Tdr absorption. Cell cycle was detected by flow cytometry. The protein levels of p21 and p27 were determined by Western blot. A549 cells was were implanted subcutaneously into nude mice to establish the xenografts tumor model, and the xenografts 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. MTT and 3H-Tdr