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该文探讨了肺癌微环境对骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BM-MSCs)自发衰老的影响及其机制研究。采用Transwell小室建立BM-MSCs与肺腺癌细胞A549非接触共培养体系,通过观察细胞形态学、细胞增殖和细胞衰老等变化以探讨肺腺癌微环境对BM-MSCs自发衰老的影响。利用qPCR技术检测细胞衰老相关端粒酶mRNA水平变化,Western blot技术寻找影响BM-MSCs自发衰老的胞内信号通路.结果显示,共培养7 d后,随着传代代次的增加,共培养组骨髓间充质干细胞(Co-MSCs)细胞形态逐渐由成纤维样漩涡状生长分化为纺锤样集落状生长,细胞增殖速度逐渐增快。同时,与BM-MSCs相比,Co-MSCs没有出现明显的衰老特征,Co-MSCs内端粒酶活性始终处于较低水平,统计学差异显著(P<0.01)。Western blot结果显示,随着传代次数的增加,BM-MSCs的信号转导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)及其磷酸化水平始终处于较低水平,而Co-MSCs的较高,且有逐渐上升的趋势,具有显著差异(P<0.05)。结果表明,肺腺癌微环境诱导Co-MSCs向着永生化细胞方向分化,有助于CoMSCs逃脱衰老,而这可能通过STAT3信号通路的持续活化来调控。
This article explored the effect of lung cancer microenvironment on spontaneous aging of bone marrow-derived mesenchymal stem cells (BM-MSCs) and its mechanism. Transwell chamber was used to establish a non-contact co-culture system of BM-MSCs and lung adenocarcinoma A549 cells. The morphological changes, cell proliferation and cell senescence were observed to investigate the effect of lung adenocarcinoma microenvironment on spontaneous aging of BM-MSCs. QPCR technique was used to detect the change of telomerase mRNA level in cell senescence and the Western blot was used to find the intracellular signaling pathway that affected the spontaneous senescence of BM-MSCs.The results showed that with the increase of subculture time, the co-culture group The morphology of BMSCs (Co-MSCs) gradually differentiated into spindle-like colony from fibroid-like swirling cells, and the cell proliferation rate increased gradually. At the same time, compared with BM-MSCs, there was no obvious aging characteristic of Co-MSCs. The telomerase activity of Co-MSCs was always at a low level, with statistical significance (P <0.01). Western blot results showed that the signal transducer and activator of transcription 3 (STAT3) and phosphorylation level of BM-MSCs were always low with the increase of passages, while Co-MSCs (P <0.05), and there was a trend of gradual increase. The results showed that the microenvironment of lung adenocarcinoma induced Co-MSCs to differentiate into immortalized cells and help CoMSCs escape from senescence, which may be regulated by the sustained activation of STAT3 signaling pathway.