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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 investigate the effects of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) -induced mesenchymal stem cells (MSCs) on chemoresistance of colon cancer cells The impact and mechanism. Methods The MSCs were treated with inflammatory cytokines IFN-γ and TNF-α. The supernatant was collected and cultured in human colon cancer cell lines HCT116 and HT29. Chemotherapy cisplatin and 5-fluorouracil were also given respectively. The morphological changes of the cells in each group were observed under the microscope. The proliferation and apoptosis of the cells were detected by MTT assay and flow cytometry. The expressions of Bax and Bcl-2 were detected by RT-PCR. Results Compared with the cells cultured without conditioned medium, the morphological changes of cells cultured in conditioned medium were more slight, the cell proliferation rate increased (P <0.05) and the apoptosis rate decreased (P <0.05) ), Bcl-2 mRNA expression was up-regulated and Bax mRNA expression was down-regulated. Conclusion MSCs induced by inflammatory cytokines IFN-γ and TNF-α can promote the chemoresistance of colon cancer cells.