摘要
目的建立人小细胞肺癌(SCLC)阿霉素(ADM)多药耐药细胞系H446/ADM模型,分析凋亡相关基因Bcl-2家族在SCLC耐药性产生中的可能作用。方法采用ADM高浓度反复间歇诱导的方法,建立人SCLC的ADM多药耐药细胞系H446/ADM模型,流式细胞术分析细胞周期变化;MTT法分析细胞系的耐药谱。流式细胞术和Westernblot法检测Bcl-2、Bcl-xL、Bak及Bax蛋白表达的变化。结果相对于亲代H446细胞,H446/ADM的生长速度减慢,细胞倍增时间延长,细胞周期分布G0/G1期细胞增加,G2/M期细胞减少;MTT法耐药谱分析示该细胞系对ADM的耐药倍数为33.09,此外,该细胞系对其他化疗药物如DNR、VCR、TAX、DDP、VP-16、MIT亦有交叉耐药。流式细胞术及Westernblot法检测示H446/ADM细胞中Bcl-2和Bcl-xL表达明显高于H446,而Bak和Bax的表达水平明显降低(P<0.05)。结论人SCLC耐药细胞系H446/ADM成功建立,有多药耐药性;Bcl-2家族蛋白表达的变化可能与耐药性产生有关。
Objective To establish an adriamycin （ADM） multi-drug resistant human small cell lung cancer cell line H446/ADM model and investigate the possible role of Bcl-2 family proteins in the producing of drug resistance. Methods H446/ADM model was obtained by exposing at intervals and repeatedly to high dose of ADM. Flow cytometry was used to analyze its cell cycle distribution. MTT assay was used to measure the cross-resistant profile of H446/ADM cell line. Expression of Bcl-2, Bcl-xL, Bak and Bax were measured by flow cytometry and Western blot in H446/ADM and H446. Re- suits H446/ADM grew more slowly and had longer population double time than its parental cell line H446. The percentage of cells decreased in G0/G1 phase increased, and decreased in G2/M phase in H446/ADM. The drug-resistance multiple of H446/ADM with regard to ADM was 33.09. H446/ADM presents cross-drug resistance not only to ADM but also to DNR, VCR, TAX, DDP, VP-16 and MIT. The expression of Bcl-2 and Bcl-xL increased while bak and bax decreased in H446/ADM compared with H446 （P 〈 0.05）. Conclusion Drug-resistant human small cell lung cancer cell line H446/ ADM can be successfully established. Bcl-2 family proteins may be involved in the producing of drug resistance.
出处
《山东医药》
CAS
北大核心
2009年第3期31-34,共4页
Shandong Medical Journal
基金
国家自然科学基金项目(30570806)
