Trichostatin A promotes GLI1 degradation and P21 expression in multiple myeloma cells
Trichostatin A promotes GLI1 degradation and P21 expression in multiple myeloma cells
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曲古抑菌素 A 促进多发性骨髓瘤细胞中 GLI1 降解和 P21 表达
DOI：
10.2147/cmar.s167330
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发表时间：
2018-01-01
影响因子：
3.3
通讯作者：
Liu, Zhiqiang
中科院分区：
文献类型：
--
作者：
Geng, Yan;Liu, Jing;Liu, Zhiqiang
背景：组蛋白去乙酰酶抑制剂是未来应用于癌症治疗的很有前途的药物。曲古抑菌素A(TSA)是一种组蛋白去乙酰化酶抑制剂，在多种癌症中显示出有效的抗肿瘤作用。然而，TSA对多发性骨髓瘤(MM)的作用及其机制尚未得到充分的研究。方法采用TSA处理的RPMI8226和MM.1S细胞进行细胞增殖、细胞周期和存活检测，分别用免疫荧光、免疫沉淀、Western blotts和qPCR方法分析GLI1蛋白和靶基因P21的定位和转录后修饰。结果TSA对MM细胞具有时间和剂量依赖性的细胞毒作用，抑制MM细胞的增殖，诱导p21蛋白表达上调，同时细胞周期蛋白D1表达降低。TSA处理导致GLI1表达下调，GLI1的核积累也受到抑制。由于Hedgehog的抑制，TSA处理后MYC和Survivin的表达明显减弱。此外，TSA以蛋白酶体依赖的方式加速GLI1的降解。此外，p21的诱导也通过减少GLI在mRNA水平的转录而下调GLI1的表达。救援实验证实，外源表达GLI1可减轻TSA诱导的MM细胞凋亡。结论TSA可抑制MM细胞生长，诱导细胞凋亡。抑制Hedgehog信号转导是TSA细胞毒作用的重要机制。
Background Histone deacetylase inhibitors are promising drugs for the future application in cancer therapy. Trichostatin A (TSA), a histone deacetylase inhibitor, exhibits effective antitumor effects in various cancers. However, the effects and underlying mechanisms of TSA on multiple myeloma (MM) are not fully investigated. Methods In the present study, RPMI8226 and MM.1S cells treated with TSA were used for cell proliferation, cell cycle, and survival examinations, then the localization and post transcriptional modification of GLI1 protein as well as the target gene P21 were analyzed using immunofluorescence, immunoprecipitation, western blots and qPCR, respectively. Results TSA exerted a time and dose-dependent cytotoxicity on MM cell lines, and suppressed the proliferation of MM cells and induced an upregulation of p21 protein accompanied by a decreased expression of cyclin D1. TSA treatment led to a downregulation of GLI1, and the nuclear accumulation of GLI1 was also inhibited. As a result of hedgehog inhibition, the expression of MYC and SURVIVIN was greatly weakened after TSA treatment. Furthermore, TSA accelerated GLI1 degradation in a proteasome-dependent manner. Additionally, p21 induction also contributed to GLI1 downregulation via reducing the transcription of GLI in mRNA level. Rescue experiments verified that exogenous expression of GLI1 alleviated MM cell apoptosis induced by TSA. Conclusion These results indicated that TSA represses MM cell growth and induces cell apoptosis. The inhibition of hedgehog signaling is an important mechanism accounting for the cytotoxic effects of TSA.