IFN-γ和TNF-α诱导间充质干细胞促进结肠癌细胞的化疗抵抗

目的 观察炎症因子干扰素γ(interferon-γ,IFN-γ)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)诱导间充质干细胞(mesenchymal stem cells,MSCs)对结肠癌细胞化疗抵抗的影响及机制。 方法 应用炎症因子IFN-γ和TNF-α联合处理MSCs,收集条件培养上清,并将其作用于人结肠癌细胞系HCT116及HT29细胞,同时分别给予化疗药物顺铂和5-氟尿嘧啶处理。显微镜下观察各组细胞形态变化,MTT法和流式细胞术检测细胞增殖和凋亡情况,RT-PCR检测凋亡相关基因Bax和Bcl-2表达情况。 结果 经化疗药物处理,与未经条件培养上清培养的细胞比较,经条件培养上清培养的细胞形态学改变更轻微,细胞增殖率增高(P<0.05)、凋亡率降低(P<0.05),Bcl-2 mRNA表达水平上调、Bax mRNA表达水平下调。 结论 经炎症因子IFN-γ和TNF-α诱导的MSCs能够促进结肠癌细胞化疗抵抗能力。
Objective To observe the influence of interferon-γ(IFN-γ)and tumor necrosis factor-α(TNF-α)treated mesenchymal stem cells (MSCs) on the chemotherapy resistance of colon cancer cells(CRCs) and to discuss the related mechanism. Methods The supernatants of MSCs treated with IFN-γ and TNF-α were collected and used, together with chemotherapy drug cisplatin and 5-fluorouracil, to treat HCT116 and HT29 CRCs. Then the cellular morphology was observed under microscope, and the cell proliferation and apoptosis were examined by MTT and PI/Annexin Ⅴ-FITC assay. Furthermore, the mRNA levels of Bax and Bcl-2 were detected by RT-PCR. Results The CRCs treated with the supernatant of MSCs exposed to inflammatory factors, compared to CRCs treated with the supernatant of MSCs not exposed to inflammatory factors, had a slighter morphology changes, a significantly higher proliferation rate (P<0.05), and a significantly lower apoptosis rate following chemotherapy(P<0.05). Moreover, Bcl-2 mRNA level was higher and Bax mRNA level was lower in CRCs treated with the supernatant of MSCs exposed to inflammatory factors. Conclusion MSCs stimulated with inflammation factors IFN-γ and TNF-α can promote the chemotherapy resistance of human CRCs