IFN-β基因修饰的人脐带源性间充质干细胞抑制A549和H226肺癌细胞的克隆、迁移和增殖能力

IFN-β gene modified human umbilical cord derived mesenchymal stem cells inhibit the colony formation, migration and proliferation of lung cancer cells A549 and H226

目的探讨干扰素-β(interferon-β,IFN-β)基因修饰的人脐带源性间充质干细胞(human umbilical cord mesenchymal stem cells,huc MSCs)对非小细胞肺癌细胞株A549和H226克隆形成、迁移和增殖能力的影响。方法按照慢病毒IFN-β感染huc MSCs条件分为以下5组:慢病毒组、阴性对照病毒组、间充质干细胞组、干扰素组、空白对照组。将IFN-β慢病毒感染huc MSC,荧光显微镜观察荧光表达,确定最佳感染复数(multiplicities of infection,MOI)值,以最佳MOI值感染huc MSCs细胞。达到稳定感染后收集慢病毒组、阴性对照病毒组和间充质干细胞组的上清,ELISA法和Western blot检测以上3组的IFN-β表达的水平。克隆形成实验、Transwell小室迁移实验、细胞增殖-毒性检测法(CKK-8)检测A549、H226细胞克隆形成、迁移和增殖能力。结果慢病毒组和阴性对照病毒组的慢病毒感染效率均在85%以上,慢病毒组的IFN-β表达水平明显高于阴性对照病毒组和间充质干细胞组,慢病毒组A549细胞和H226细胞的克隆形成、迁移能力和增值能力明显降低。结论 IFN-β修饰的huc MSCs能显著抑制非小细胞肺癌细胞株A549和H226的克隆形成、迁移及增值能力,有望成为治疗非小细胞肺癌的一种新途径。

Objective To investigate the effects of interferon-β（IFN-β） gene modified human umbilical cord derived mesenchymal stem cells（huc MSCs） on the colony formation, migration and proliferation of non-small cell lung cancer（NSCLC） cell line A549 and H226. Methods The optimal multiplicity of infection（MOI） for lentivirus infection was determined in huc MSCs. NSCLC cell line A549 and H226 were exposed to the conditioned media collected from huc MSCs that were transduced with lentivirus-IFN-β-RFP, control lentivirus or none. For comparison, A549 and H226 were also treated with regular culture medium（blank control） or IFN-β（positive control）. The levels of IFN-β protein in the medium were analyzed by Human IFN-β ELISA Kit and Western blotting. The impacts of conditioned media on cell colony formation, migration and proliferation were evaluated through low-density cell colony formation, Transwell migration assay and CCK-8 proliferation assay respectively. Results The virus infection efficiency was greater than 85% in both virus transduced groups, and IFN-β expression level in lentivirus-IFN-β-RFP group was significantly higher than in the controls. Compared to blank medium control, IFN-β and conditioned medium from lentivirus-IFN-β-RFP infected huc MSCs could significantly inhibit the colony formation, migration and proliferation were significantly decreased in A549 and H226 cells. Conclusion IFN-β, synthetic or secreted by huc MSCs, can effectively inhibit the colony formation, migration and proliferation of non-small cell lung cancer cell lines A549 and H226, which may provide a new therapeutic path for non-small cell lung cancer.