博来霉素水解酶是预测博来霉素敏感性的分子标志物
首发时间:2016-05-20
摘要:目的:研究博来霉素水解酶(BLH)是否为肿瘤细胞对博来霉素敏感性的生物标志物。方法: MTT法检测细胞的存活率变化,Western bloting检测相关分子的蛋白表达, RNA干扰技术降低细胞内BLH蛋白的表达,流式细胞仪检测细胞周期和活性氧自由基的变化。 结果: 通过8种细胞筛选,确定BLH表达较高的HeLa细胞进行深入研究。使用RNA干扰方法降低BLH蛋白的表达,明显增加了对博来霉素的敏感性,使活性氧自由基水平升高,阻断在G2/M期的细胞增多。使用非致死浓度的博来霉素和多柔比星处理,引起BLH表达明显增加。结论:BLH是预测博来霉素敏感性的生物标志物,可能是一种应激性蛋白。
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Bleomycin hydrolase is a Biomarker to predict the Susceptibility to Bleomycin
Abstract:Objective: To investigate whether bleomycin hydrolase (BLH) is a biomarker for determination of susceptibility to bleomycin. Methods: Cell viability was assessed using MTT method and the protein expression was detected by western blot. The protein level of BLH was knocked down by RNA interference. Cell cycle and generation of reactive oxygen species were assayed with flow cytometer. Results: HeLa cell line was selected from 8 types of cell lines for the following experiments. The action of bleomycin was potentiated after the cellular BLH level was knockdown by RNA interference. The generation of reactive oxygen species and retardation of G2/M phase were increased by knockdown of BLH. The expression level of BLH was obviously increased after the cells were treated bleomycin or doxorucin at sublethal concentrations. Conclusion: BLH is a a biomarker to predict the susceptibility to bleomycin and it may be play a role to stress response.
Keywords: bleomycin hydrolase Bleomycin Drug susceptibility Biomaker
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