Tanshinone IIA inhibits human prostate cancer cells growth by induction of endoplasmic reticulum stress in vitro and in vivo
Tanshinone IIA inhibits human prostate cancer cells growth by induction of endoplasmic reticulum stress in vitro and in vivo
复制标题
DOI：
10.1038/pcan.2013.38
复制
发表时间：
2013-12-01
影响因子：
4.8
通讯作者：
Pang, C. Y.
中科院分区：
文献类型：
--
作者：
Chiu, S. C.;Huang, S. Y.;Pang, C. Y.
背景：丹参酮IIA （Tan-IIA）是从丹参根中分离得到的主要亲脂性成分之一。我们在体外和体内研究了Tan-IIA治疗前列腺癌细胞诱导细胞死亡的机制。方法：用Tan-IIA处理细胞，观察细胞生长抑制情况。流式细胞术检测Tan-IIA处理后的细胞周期谱。Tan-IIA处理后，检测细胞周期调节蛋白和凋亡相关蛋白的表达水平。测定内质网（ER）应激调节基因的表达水平，探讨其在tan - iia诱导的细胞死亡中的作用。转染小干扰RNA （siRNA）可抑制GADD153的表达。用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑测定细胞的死亡率和增殖率。在LNCaP异种移植瘤模型中检测Tan-IIA的抗肿瘤活性。结果：我们的研究结果显示，Tan-IIA以剂量依赖的方式导致前列腺癌细胞死亡，并且在LNCaP细胞中注意到细胞周期阻滞在G0/G1期。G0/G1期阻滞与CDK抑制剂（p16、p21和p27）水平升高和检查点蛋白水平降低相关。Tan-IIA还诱导前列腺癌细胞内质网应激：鉴定出GADD153/CCAAT/增强子结合蛋白同源蛋白（CHOP）的活化和核易位，并证实下游分子GRP78/BiP、肌醇需要蛋白-1 α和GADD153/CHOP的表达增加。通过siRNA阻断GADD153/CHOP表达可减少tania诱导的LNCaP细胞死亡。Tan-IIA还抑制LNCaP异种移植肿瘤的生长，治疗13天后肿瘤体积减少86.4%。结论：我们的研究结果表明，Tan-IIA导致LNCaP细胞的G0/G1细胞周期阻滞，其细胞毒性至少部分是由内质网应激诱导介导的。这些数据为支持Tan-IIA在前列腺癌中通过诱导内质网应激作为潜在的抗癌药物提供了证据。
BACKGROUND: Tanshinone IIA (Tan-IIA) is one of the major lipophilic components isolated from the root of Salviae Miltiorrhizae Radix. We explored the mechanisms of cell death induced by Tan-IIA treatment in prostate cancer cells in vitro and in vivo.METHODS: Cells were treated with Tan-IIA and growth inhibition was assessed. Cell cycle profiles after Tan-IIA treatment were determined by flow cytometry. Expression levels of cell cycle regulatory proteins and apoptosis-related proteins were determined after Tan-IIA treatment. Expression levels of endoplasmic reticulum (ER) stress-regulated genes were determined to investigate their role in Tan-IIA-induced cell death. GADD153 expression was knocked down by small interfering RNA (siRNA) transfection. Rate of cell death and proliferation was obtained by 3-(4,5-dimethyl thizol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Antitumor activity of Tan-IIA was performed in LNCaP xenograft model.RESULTS: Our results showed that Tan-IIA caused prostate cancer cell death in a dose-dependent manner, and cell cycle arrest at G0/G1 phase was noted, in LNCaP cells. The G0/G1 phase arrest correlated with increase levels of CDK inhibitors (p16, p21 and p27) and decrease of the checkpoint proteins. Tan-IIA also induced ER stress in prostate cancer cells: activation and nuclear translocation of GADD153/CCAAT/enhancer-binding protein-homologous protein (CHOP) were identified, and increased expression of the downstream molecules GRP78/BiP, inositol-requiring protein-1 alpha and GADD153/CHOP were evidenced. Blockage of GADD153/CHOP expression by siRNA reduced Tan-IIA-induced cell death in LNCaP cells. Tan-IIA also suppressed LNCaP xenograft tumor growth, causing 86.4% reduction in tumor volume after 13 days of treatment.CONCLUSIONS: Our findings suggest that Tan-IIA causes G0/G1 cell cycle arrest in LNCaP cells and its cytotoxicity is mediated at least partly by ER stress induction. These data provide evidence supporting Tan-IIA as a potential anticancer agent by inducing ER stress in prostate cancer.