慢病毒介导的PIM-1基因调控大鼠骨髓间充质干细胞增殖和对凋亡的影响

Effect on proliferation and apoptosis of bone marrow mesenchymal stem cells regulated by PIM-1 gene mediated by lentivirus

目的:观察PIM-1基因调控大鼠骨髓间充质干细胞(BMMSCs)的增殖和对凋亡的影响。方法:应用流式细胞仪(FCM)分别对第4代(P4)BMMSCs表面抗原CD29、CD44和CD45进行鉴定。实验分为空白对照组(对照组)、EGFP转染组(EGFP组)、PIM-1基因转染组(PIM-1组),将成功构建的慢病毒载体LV-egfp-PIM-1及LV-egfp转染至BMMSCs中,采用PCR法检测慢病毒载体转染后PIM-1表达的变化,用Western blot法检测慢病毒载体转染后PIM-1蛋白表达水平,用MTT比色法检测PIM-1对BMMSCs增殖能力的调控作用,FCM检测PIM-1对BMMSCs凋亡的影响。结果:大鼠BMMSCs成功转入PIM-1基因后,与对照组相比,PIM-1蛋白的表达显著增加(P<0.01),细胞增殖能力显著增加(P<0.01),细胞凋亡率则显著降低(P<0.01)。结论:过表达PIM-1可显著提高BMMSCs的增殖能力和抑制BMMSCs凋亡。

AIM： To explore the effect on the proliferation and the apoptosis of bone marrow mesenchymal stem ceils （BMMSCs） regulated by PIM-1 gene mediated by lentivirus. METHODS： Antigen expressions of CD29, CD44, and CD45 were assayed in the ceils of the fourth passage by flow cytometry to identify BMMSCs. The experiment was divided into control group, EGFP transfected group （EGFP group）, and PIM-1 gene transfection group （PIM-1 group）. The combinant lentivirus vector-PIM-1 （Lvefgp-PIM-1 ） and lentivirus vector-EGFP were transfected into BMMSCs. PCR was performed to detect the expression of PIM-1. Western blot was used to detect the expression of PIM-1 protein level. MTT was used to evaluate the proliferation of BMMSCs regulated by PIM-1, and fluorescence-activated cell sorting （FACS） technology was employed to detect the rate of cell apoptosis. RESULTS： PIM-1 was successfully transfected into BMMSCs. Compared with those in control group, the cell PIM-1 protein levels and the cell proliferation activity significantly increased （both P 〈0. 01 ）, whereas the cell apoptosis rate significantly decreased （P 〈 0. 01 ）. CONCLUSION ： LV-PIM-1 transfection can markedly promote BMMSC growth and inhibit cell apoptosis.