解剖学报

解剖学报 ›› 2016, Vol. 47 ›› Issue (6): 774-778.doi: 10.16098/j.issn.0529-1356.2016.06.009

• 肿瘤生物学 • 上一篇    下一篇

雷公藤甲素通过诱导细胞自噬促进结肠癌CT26细胞死亡

张天娇 韩森 张玮 赵福建 李健*   

  1. 北京中医药大学基础医学院组织学胚胎学教研室，北京 100029
  • 收稿日期:2016-04-26 修回日期:2016-07-08 出版日期:2016-12-06 发布日期:2016-12-06
  • 通讯作者: 李 健 E-mail:ljbeijing1970@sina.com.cn
  • 基金资助:

    从FXR/PXR/CYP3A4信号通路探讨雷公藤诱发胆汁淤积性肝损伤的分子

Triptolide promoting colon cancer CT26 cell death through induction of autophagy

ZHANG Tian-jiao HAN Sen ZHANG Wei ZHAO Fu-jian LI Jian*   

  1. Department of Histology and Embryology, School of Basic Medical Science, Beijing University of Chinese Medicine, Beijing 100029, China
  • Received:2016-04-26 Revised:2016-07-08 Online:2016-12-06 Published:2016-12-06
  • Contact: LI Jian E-mail:ljbeijing1970@sina.com.cn

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摘要：

目的 探讨雷公藤甲素诱导结肠癌CT26细胞自噬与细胞死亡之间的关系。 方法 体外培养小鼠结肠癌CT26细胞，应用免疫荧光、荧光双标自噬腺病毒载体（Ad-mRFP-GFP-LC3）技术，在激光扫描共焦显微镜下观察LC3的表达轮廓及水平，并结合免疫印迹法对LC3蛋白、P62蛋白做半定量分析；进而用单溶液细胞增殖检测（MTS）试剂测定CT26细胞增殖抑制率；用AnnexinV-FITC/PI双染、流式细胞术检测细胞凋亡/死亡率。 结果 雷公藤甲素能诱导CT26细胞出现明确的自噬流；激活细胞自噬可提高CT26细胞的晚期凋亡率。与雷公藤甲素组比较，雷帕霉素与其联用能显著增强CT26细胞死亡。 结论 自噬可能介导了雷公藤甲素诱导CT26细胞死亡的过程。

关键词: 雷公藤甲素, CT-26, 结肠癌, 自噬, 免疫荧光, 免疫印迹法, 流式细胞术, 小鼠

Abstract:

Objective To investigate the relationship of autophagy and cell death in CT26, a cell line from mouse colon cancer was used and treated by triptolide. Methods Immunofluorescence staining, LC3 adenovirus vector (mRFP-GFP-LC3), and Western blotting methods were adopted to examine autophagy flux in CT26 cells. The AQueous one solution cell proliferation assay(MTS) and AnnexinV-FITC/PI reagents were used to evaluate the cells inhibitions rate, apoptosis and cell death rate of CT26 cells after treated by triptolide, rapamycin, and triptolide combined with rapamycin. Results The CT26 cells showed significant autophagy flux induced by triptolide and activating cell autophagy improved the late CT26 cell apoptosis rate. Compared with the group of triptolide, the group of triptolide combined with rapamycin obviously increased CT26 cell death. Conclusion Triptolide promotes colon cancer CT26 cell death through induction of autophagy.

Key words: Triptolide, CT-26, Colon cancer, Autophagy, Immunofluorescence, Western blotting, Flow cytometry, Mouse