Triptolide induces apoptotic cell death of human cholangiocarcinoma cells through inhibition of myeloid cell leukemia-1

BMC Cancer

Xiwei Ding¹  Bin Zhang¹  Qingshan Pei³  Jianmei Pan²  Shuling Huang¹  Yan Yang¹  Zhu Zhu¹  Ying Lv¹  Xiaoping Zou¹ 

[1] Department of Gastroenterology, Drum Tower Hospital, Affiliated to Medical School of Nanjing University, Nanjing, China

[2] Department of Gastroenterology, Jinan Central Hospital, Affiliated to Shandong University, Jinan, China

[3] Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, China

关键词: Apoptosis;    Mcl-1;    Cholangiocarcinoma;    Triptolide;   

Others  :  858886
DOI  :  10.1186/1471-2407-14-271

 received in 2013-12-14, accepted in 2014-04-08,  发布年份 2014

【 摘 要 】

Background

Cholangiocarcinoma (CCA), a devastating neoplasm, is highly resistant to current chemotherapies. CCA cells frequently overexpress the antiapoptotic protein myeloid cell leukemia-1(Mcl-1), which is responsible for its extraordinary ability to evade cell death. Triptolide, a bioactive ingredient extracted from Chinese medicinal plant, has been shown to inhibit cell proliferation and induce apoptosis in several cancers.

Methods

CCK-8 assay was performed to detect cell survival rate in vitro. DAPI staining and Flow cytometry were used to analyze apoptosis. Western blot was performed to determine the expression levels of caspase-3, caspase-7, caspase-9, PARP, and Mcl-1. Quantitative real-time PCR and immunofluorescence were used to detect the expression levels of Mcl-1. The nude mice xenograft model was used to evaluate the antitumor effect of triptolide in vivo.

Results

Triptolide reduced cell viability in cholangiocarcinoma cell lines in a dose- and time-dependent manner, with IC₅₀ values of 12.6 ± 0.6 nM, 20.5 ± 4.2 nM, and 18.5 ± 0.7 nM at 48 h for HuCCT1, QBC939, and FRH0201 respectively. Triptolide induced apoptosis in CCA cell lines in part through mitochondrial pathway. Using quantitative real-time PCR, western blot and immunofluorescence, we have shown that triptolide downregulates Mcl-1 mRNA and protein levels. Furthermore, triptolide inhibited the CCA growth in vivo.

Conclusions

Triptolide has profound antitumor effect on CCA, probably by inducing apoptosis through inhibition of Mcl-1. Triptolide would be a promising therapeutic agent for CCA.

【 授权许可】

   
2014 Ding et al.; licensee BioMed Central Ltd.

【 图 表 】

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