基础医学与临床 ›› 2011, Vol. 31 ›› Issue (7): 814-819.

• 研究论文 • 上一篇    下一篇

自噬增强结肠癌CD133+细胞对5-FU的化疗抵抗

司晓丽, 刘伟, 杨爱军, 王晨昱, 李敏   

  1. 兰州大学基础医学院 病理学研究所,甘肃省新药临床前研究重点实验室,甘肃 兰州730000
  • 收稿日期:2010-11-24 修回日期:2011-01-03 出版日期:2011-07-05 发布日期:2011-07-05
  • 通讯作者: 李敏 E-mail:Limin@lzu.edu.cn

Autophagy Enhances Chemotheragy Resistance of CD133+ Colon Cancer Stem Cells to 5-FU

SI Xiao-li, LIU Wei, YANG Ai-jun, WANG Chen-yu, LI Min*   

  1. Institute of Pathology, College of Basic Medicine, Lanzhou University, Gansu Key Laboratory for Preclinical Study of New Drugs, Lanzhou 730000, China
  • Received:2010-11-24 Revised:2011-01-03 Online:2011-07-05 Published:2011-07-05
  • Contact: Min LI E-mail:Limin@lzu.edu.cn

摘要: 目的 对5-FU诱导结肠癌CD133+细胞发生自噬的过程进行观察,初步探讨自噬对化疗的影响。 方法 用MTT法检测细胞增殖抑制率及IC50;用免疫磁珠分选CD133+细胞;用透射电子显微镜(TEM)观察细胞的超微结构,用MDC染色,荧光显微镜观察自噬囊泡;联合应用细胞活性实验、集落形成实验研究自噬对化疗的影响。结果 不同浓度5-FU处理后,人结肠癌SW480细胞活性明显受到抑制,其半数抑制浓度IC50为(1.09±0.18)μg/mL。CD133+胞质出现大量的自噬体及自噬囊泡; 相比较单独给予5-FU,联合给予5-FU及3-MA,CD133+细胞活性由(81.8±4.6)%下降到(56.3±5.5)%(P<0.05), 集落形成率由(81.1±3.4)%下降(64.4±4.8)%(P<0.05)。结论 5-FU可诱导结肠癌CD133+细胞发生自噬;自噬的抑制能够增强5-FU对结肠癌CD133+细胞的细胞毒效应。

Abstract: Objective To observe the autophagy of CD133+ CSCs induced by 5-FU,and explore the effect of autophagy on the chemotherapy Methods The cell proliferative inhibition rates and IC50 of 5-FU on human colon cancer cell lines SW480 were measured by MTT assay. CD133+ CSCs were purified with MACS(magnetic activated cell-sorting system) CD133 cell isolation kit. Then the morphplpgical features of cells were observed using transmission election microscopy(TEM),and the autophagic vacuoles(AV) was observed using fluorescent microscope by monodansylcadaverin(MDC) staining. The effect of autophagy on the chemotherapey was evaluated by cell viability assay and colony-forming test. Results The vibility of human colon cancer cell line SW480 was obviously inhibited with different contrations of 5-FU,half of inhibition concentration(IC50) value was (1.09±0.18)μg/mL .Autophagsomes and autophagy vesica were observed in CD133+ CSCs. The combination treatment of 5-FU and 3-MA significantly decreased the viability of CD133+CSCs from (81.8±4.6)% to (56.3±5.5)%(P<0.05), and the colony-forming efficiency from (81.1±3.4)% to (64.4±4.8)% (P<0.05), compared with 5-FU alone. Conclusion 5-FU may induce autophagy in CD133+ CSCs. Inhibition of autophagy may be employed to increase the cytotoxic effect of CD133+ CSCs to 5-FU.

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