慢病毒介导IL2基因感染MSCs对胰腺癌PANC-1治疗效应

The Therapeutic Effect of IL-2 Expressed MSCs in the Treatment of Pancreatic Cancer Cell Line PANC-1 in Vitro

为了评价慢病毒介导IL2基因感染间充质干细胞(MSCs)对胰腺癌PANC-1增殖和迁移体外模型中的效应,本研究在体外分离、培养和鉴定了MSCs后,采用Gateway技术构建IL-2表达载体,采用HEK293细胞产生慢病毒颗粒,感染细胞后采用免疫印迹法进行鉴定。通过Transwll小室实验,采用IL-2/MSCs进行处理后,观察肿瘤的凋亡情况。此外,本研究还采用Caspase抑制剂及IL-2单抗对IL2/MSC促肿瘤凋亡的机制进行明确。研究结果表明,成功分离的间充质干细胞,经流式鉴定后为CD34、CD45不表达,CD29、CD44和CD90高表达。体外实验的结果显示IL-2/MSCs能够显著增加肿瘤的凋亡。IL-2/MSCs能够通过活化Casapase相关的信号发挥抗凋亡效应。研究结论初步表明,IL-2/MSC具有一定的胰腺癌治疗效应,其主要是通过活化Caspase来发挥对肿瘤的杀伤作用。

The aim of this study was to evaluate the therapeutic effect of IL-2 expressed mesenchymal stem cells(MSCs) in the treatment of pancreatic cancer cell line PANC-1 in vitro. MSCs were isolated, cultured and characterized in vitro. The IL-2 expression vector was constructed by Gateway technology and packaged into HEK293 cell to produce the lentivirus. The infected MSCs were characterized by Western blotting. We also established transwell system and treated with IL-2/MSCs to evaluate the therapeutic effect on tumor cell apoptosis.MSCs were successfully isolated and the flow cytometry showed MSCs were expressed CD29, CD44 and CD90,but not CD34 and CD45. The in vitro data showed that IL-2/MSCs could significantly promote the tumor cell apoptosis. The apoptosis effect was mediated by caspase signaling system. The conclusion of this study showed that IL-2/MSCs could exert therapeutic effect on pancreatic cancer through activating caspase system.