PLAUR基因过表达对人结肠癌细胞株CD133^+、CD44^+细胞增殖的影响

Effect of the overexpression of PLAUR gene on the proliferation of CD133^+ & CD44^+ human colon cancer cell lines

目的探讨尿激酶型纤溶酶原激活因子受体(urokinase-type plasminogen activator receptor,PLAUR)过表达对人结肠癌细胞株CT-26中CD133+、CD44+细胞增殖的影响。方法用携带PLAUR基因的逆转录病毒载体(pMSCV-PLAUR)感染人结肠癌CT-26/CD133+以及CT-26/CD44+细胞,作为观察组(pMSCV-PLAUR/CD133+组,pMSCV-PLAUR/CD44+组);以未携带PLAUR基因的逆转录病毒载体(pMSCV)感染CT-26/CD133+、CT-26/CD44+细胞,作为对照组(pMSCV/CD133+组,pMSCV/CD44+组)。用实时定量荧光PCR和免疫印迹法分别检测细胞中PLAUR mRNA水平和蛋白表达的水平;利用平板克隆实验检测细胞增殖能力的变化。结果在观察组和对照组中PLAUR mRNA的表达情况分别为CD133+(1.02±0.20)、(0.52±0.12)和CD44+(1.15±0.09)、(0.68±0.07),观察组PLAUR蛋白表达的水平均较对照组显著增高(均P<0.05);观察组、对照组的细胞克隆形成率分别为CD133+(78.25±3.29)%,(51.45±2.53)%以及CD44+(82.45±4.56)%,(44.96±5.64)%,观察组高于对照组(P<0.05)。结论PLAUR过表达可以显著增强人结直肠癌CT-26/CD133+、CT-26/CD44+细胞的克隆形成能力,促进CD133+、CD44+细胞增殖。

Objective To investigate the effect of the overexpression of urokinase type plasminogen activator receptor(PLAUR)on the proliferation of CD133+&CD44+cells in human colon cancer cell line CT-26.Methods Human colon cancer CT-26/CD133+and CT-26/CD44+cells were infected with retroviral vector carrying PLAUR gene(pMSV-PLAUR/CD133+group,pMSV-PLAUR/CD44+group),and CT-26/CD44+cells were infected with retroviral vector without PLAUR gene(pMSV/CD133+group,pMSV/CD44+group).The level of PLAUR mRNA and protein expression were detected by real-time quantitative RT-PCR and Western blotting,respectively,and the cell proliferation was detected by plate cloning.Results The expression of PLAUR mRNA was(1.02±0.20),(0.52±0.12),(1.15±0.09)and(0.68±0.07)in the observation group and the control group,respectively.The expression of PLAUR mRNA and protein in the observation group was significantly higher than that in the control group(P<0.05),and the cell clone formation rates in the observation group and the control group were(78.25±3.29)%,(51.45±2.53)%,(82.45±4.56)%and(44.96±5.64)%,the observation group was higher than the control group(P<0.05).Conclusion The overexpression of PLAUR can significantly enhance the clonogenic ability of CT-26/CD133+,CT-26/CD44+cells and promote the proliferation of CD133+,CD44+cells.