长期缺氧对肺腺癌 SPCA1细胞谷胱甘肽转移酶表达及耐药性的影响  

Prolonged hypoxia induces GST-π expression and drug resistance in lung adenocarcinoma cell line SPCA1

作　　者：柳永蕾[1] 宋现让[1] 王兴武[1] 魏玲[1] 李大鹏[1] 

机构地区：[1]山东省肿瘤医院基础研究中心,济南250117

出　　处：《中华结核和呼吸杂志》2007年第2期108-111,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金(30570547)

摘　　要：目的探讨长期缺氧对 SPCA1细胞谷胱甘肽转移酶(GST-π)表达的影响及耐药性的改变。方法 SPCA1细胞常氧(20%O_2)和缺氧(0.5%O_2)条件下作用16 h 后,提取 RNA 和蛋白,用定量 RT-PCR 的方法检测缺氧诱导因子1α(HIF-1α)和GST-π mRNA 的表达情况,用 Western blotting法检测 HIF-1α蛋白;制备细胞悬液,用流式细胞仪测定 GST-π表达;通过克隆形成实验分析 SPCA1细胞对阿霉素和丝裂霉素的药物敏感性。通过 RNA 干扰技术下调 HIF-1α表达后,分为对照组和干扰组,观察 GST-π的表达。结果设常氧组 GST-π mRNA 表达率为1,缺氧组为12.2。常氧组 GST-π蛋白阳性表达率为(35.78±1.25)%,缺氧组为(72.13±2.11)%。与常氧组比较缺氧组 HIF-1α表达明显增加。计算1%克隆存活的药物浓度,阿霉素处理细胞,常氧组浓度为(0.29±0.05)μg/ml;缺氧组浓度为(0.48±0.07)μg/ml;丝裂霉素处理细胞,常氧组浓度为(0.71±0.10)μg/ml;缺氧组浓度为(0.79±0.12)μg/ml。干扰 HIF-1α后,HIF-1α mRNA 与对照组比较下降了70%,而 GST-π mRNA稍有下降。GST-π蛋白表达常氧条件下对照组为(32.14±1.23)%,干扰组为(30.88±1.65)%;缺氧条件下对照组为(64.78±2.45)%,干扰组为(67.42±2.21)%。结论长期缺氧可诱导 SPCA1细胞HIF-1α和 GST-π表达增加,对阿霉素耐药性增加,对丝裂霉素没有影响,HIF-1α和 GST-π并没有直接的相关性。Objective To investigate the association of hypoxia with GST-π expression in lung adenocarcinoma cells SPCA1 and the drug resistance in hypoxic condition. Methods RNA and protein in SPCA1 cells treated with hypoxia （0. 5%O2） for 16 h were isolated. HIF-1α and GST-π mRNA expression was detected by real time RT-PCR, and HIF-1α protein was detected by Western blotting analysis. GST-π protein was finally analyzed on a FACScan flow cytometer using Cellquest software. Sensitivity of Doxorubicin and Mitomycin in hypoxic SPCA1 cells was detected using cell clonogenic assay. Results HIF-1α expression was up-regnlated on mRNA and protein levels in hypoxic SPCA1 cells, so as was GST-π expression. Doxorubicin resistance was increased, but hypoxia had no significant influence on Mitomycin resistance. GST-π expression had no significant change after HIF-1α was down-regulated by RNA interfering. Conclusion Though prolonged hypoxia can induce GST-π expression and increase drug resistance such as to Doxorubicin, HIF-1α expression has no correlation with GST-π expression.

关 键 词：肺肿瘤 细胞低氧 谷胱甘肽转移酶 

分 类 号：R734.2[医药卫生—肿瘤]