白藜芦醇抑制结肠癌干细胞增殖并增强MICA/B的表达

Resveratrol inhibits cell proliferation and up-regulates MICA/B expression in human colon cancer stem cells

目的研究白藜芦醇(Res)对结肠癌干细胞(CCSC)增殖、凋亡和免疫原性的影响。方法采用无血清培养法由HCT116结肠癌细胞诱导培养CCSC,通过检测CCSC标志物CD133、CD44进行鉴定。MTT法检测Res对CCSC增殖的影响;annexinⅤ-FITC/PI双染色结合流式细胞术检测Res对CCSC凋亡的影响;流式细胞术检测Res对CCSC生长周期及主要组织相容性复合体Ⅰ类分子相关链A/B(MICA/B)表达的影响。结果 HCT116细胞在无血清培养下成球生长。球形细胞中CD133+细胞占91.07%±1.79%,CD44+细胞占90.33%±1.78%。与对照组相比,Res能明显抑制CCSC增殖,并且呈时间、剂量依赖性;Res作用48 h后,CCSC细胞周期G0/G1期比例显著升高,S期下降,呈剂量依赖性;随着药物浓度增加,CCSC的凋亡率增加,MICA/B的表达增强。结论从HCT116结肠癌细胞成功诱导培养出CCSC,Res能剂量依赖性地抑制CCSC的增殖,与阻滞细胞于G0/G1期,并诱导细胞凋亡有关;Res能增强CCSC中MICA/B的表达,增强其免疫原性。

Objective To investigate the effect of resveratrol （Res） on the proliferation, apoptosis and immunogenicity of colorectal cancer stem cells （CCSCs）. Methods The CCSCs were induced from colon cancer cell line HCT116 in serum-free medium （SFM）. The expressions of CD133 and CD44 were detected by flow cytometry to identify CCSCs. After treatment with Res at （12.5 -200.0） pmol/L, the effect of Res on CCSC proliferation was detected by MTT assay; cell apoptosis was examined by flow cytometry corrbined with annexin V-FITC/PI staining; cell cycle and the expression of major histocompatibility comp4ex class I-related chain A and B （MICA/B） were assessed by flow cytometry. Results HCTll6 cells formed cancer stem cell spheres in SFM. The proportion of CD133^＋ cells in cell spheres was （91.07 ± 1.79）%, and CD44^＋ cells was （90.33 ± 1.78）%. Compared with control groups, Res significantly inhibited CCSC proliferation in a time- and dose-depended manner. After treatment with Res for 48 hours, the proportion of cells increased in the G0/G1 phase an.d decreased in the S phase, both in a dose-depended manner. Apoptosis rate of CCSCs and the expression of MICA/B were raised with the increasing concentration of Res. Conclusion CCSCs were successfully induced from HCT116 colon cancer cell line. Res could depress CCSC proliferation in a time-and dose-depended manner, arrest cell cycle in the GO/G1 phase and promote cell apoptosis. Res could up-regulate the expression of MICA/B in CCSCs and enhance cell immunogenicity.