CD44、CD133耦连miR-425装载纳米脂质体对结直肠癌干细胞PDL-1表达的下调作用

摘要

目的 探究CD44、CD133耦连miR-425装载纳米脂质体对结直肠癌干细胞PDL-1表达的下调作用.方法选择结直肠癌细胞系HCT116分选出CD44、CD133/1+细胞,进行miR-425装载纳米脂质体转染,分为对照组(转染con-trol载体)、实验组(转染miR-425慢病毒载体),空白组以纳米脂质体代替,测定干细胞生长状态,检测PDL-1表达水平.结果分选的CD 133/1+细胞呈圆形,12 h后开始贴壁生长,状态良好.qRT-PCR检测显示实验组的miR-425的相对表达水平为(182.44±22.19),对照组与空白组分别为(1.84±1.11)和(1.67±0.98),实验组明显高于对照组与空白组(P<0.05).MTT检测结果表明,与空白组(0.78±0.22)、对照组(0.81±0.17)比较,实验组(0.24±0.15)的HCT116细胞生长速度明显减慢(P<0.05).空白组与对照组的细胞增殖率分别为(0.48±0.14)与(0.51±0.18),都明显高于实验组的(0.30±0.11).Western blot检测分析显示实验组、对照组、空白组的PDL-1相对表达量分别为(0.33±0.13)、(0.76±0.21)和(0.82±0.15),实验组明显低于对照组与空白组(P<0.05).结论CD44、CD133耦连miR-425装载纳米脂质体能抑制结直肠癌干细胞PDL-1的表达,从而显著抑制干细胞的生长、增殖能力.%Objective To investigate the PDL-1 down regulated role of CD44,CD133 coupled with miR-425 loaded nano liposome in the colorectal cancer stem cells. Methods The colorectal cancer cell line HCT116 were selected for CD44,CD133/1+ cells,and were given the miR-425 loaded nano liposome transfection,the cells were divided into the control group ( transfect-ed with control vector) and the experimental group (transfected with miR-425lentivirus vector),the blank group was the nano li-posome,and detected the cell growth and the expression level of PDL-1. Results The CD 133/1+ cells were round and grew well after 12 h. qRT-PCR test showed that the relative expression level of miR-425 in the experimental group was (182. 44 ± 22. 19),while the control group and the blank group were (1. 84 ± 1. 11) and (1. 67 ± 0. 98) respectively,the experimental group were higher than that of the control group and the blank group (P<0. 05). The results of MTT showed that compared with the blank group (0. 78 ± 0. 22) and the control group (0. 81 ± 0. 17),the growth rate of HCT116 cells (0. 24 ± 0. 15) were sig-nificantly slower in the experimental group (P<0. 05). Compared with the experimental group,the cell proliferation rate of the blank group(0. 48 ± 0. 14)and the control group(0. 51 ± 0. 18)were significantly higher than that of the experimental group(0. 30 ± 0. 11). Western blot analysis showed that the relative expression of PDL-1 in the experimental group,the control group and the blank group was 0. 33 ± 0. 13,0. 76 ± 0. 21 and 0. 82 ± 0. 15 respectively,the experimental group were significantly lower than the control group and the blank group (P<0. 05). Conclusion CD44 and CD133 coupled with miR-425 can inhibit the expression of PDL-1 in colorectal cancer stem cells,which can significantly inhibit the growth and proliferation of stem cells.