摘要
目的研究人肺腺癌细胞系A549和其顺铂耐药细胞系A549/DDP中GSK-3β的磷酸化和胞内分布以探讨GSK-3β在顺铂耐药中的作用。方法蛋白质免疫印迹法检测A549/DDP和A549细胞质和细胞核中总GSK-3β、p-GSK-3βser9和p-GSK-3βtyr6的表达。MTT法、流式细胞术分别检测顺铂耐药性、肺癌细胞凋亡率。结果 A549/DDP细胞胞质中p-GSK-3βser9水平明显高于A549细胞(P<0.01),顺铂的处理增加了A549/DDP细胞中p-GSK-3βser9的水平(P<0.01),相反却减少了A549细胞中p-GSK-3βser9的水平(P<0.01)。A549/DDP细胞质中p-GSK-3βtyr6水平明显低于A549细胞(P<0.01),顺铂的处理减少了A549/DDP细胞中p-GSK-3βtyr6的水平(P<0.01),却增加了A549细胞中p-GSK-3βtyr6的水平(P<0.01)。结论细胞质中GSK-3β活性受抑可能是非小细胞肺癌顺铂耐药的原因。
Objective To explore the role of GSK-3β in cisplatin resistance by evaluating the phosphorylation status and intracellular localization of GSK-3β in lung adenocarcinoma ceil A549 and its cisplatin resistant cell clone A549/DDP. Methods Both total and nuclear/cytoplasmic GSK-3β, p-GSK-3βser9 and p-GSK-3βtr6expression were detected by Western blot. Cispatin resistance and cell apoptotic rate were measured by MTT and flow cytometry in both A549 and A549/DDP cells. Results Increased cytoplasmic p-GSK-3βer9 was observed in A549/DDP cells as compared to A549 cells（P〈0.01）. Cisplatin increased cytoplasmic p-GSK-3βser9 in A549/DDP cells but decreased it in A549 cells（P〈0.01）. Conversely, cytoplasmic levels of p-GSK-3βtyr216 were significantly decreased in A549/DDP cells as compared to A549 cells. The levels of p-GSK-3βtyr216 in the cytoplasm of A549/DDP cells decreased with cisplatin treatment while they increased in A549 cells（P〈0.01）. Conclusion Inhibition of cytoplasmic activity of GSK-3β was responsible for cisplatin resistance in lung adenocarcinoma.
出处
《中华临床医师杂志（电子版）》
CAS
2013年第20期81-84,共4页
Chinese Journal of Clinicians(Electronic Edition)
基金
辽宁省"百千万人才工程"培养项目(2012921032)
辽宁省自然科学基金(201202285)
关键词
肺肿瘤
化疗耐药
GSK-3Β
Lung neoplasms
Chemotherapy resistance
GSK-3β
