白血病相关蛋白16调控Wnt/β-链蛋白通路对结肠癌细胞增殖、侵袭的影响

Effects of leukemia related protein 16 on Wnt/β-catenin pathway on proliferation and invasion of colon cancer cells

目的研究人白血病相关蛋白16(LRP16)调控Wnt/β-链蛋白(β-catenin)通路对结肠癌细胞增殖、侵袭的影响。方法将结肠癌HCT116细胞分为3组:空白组、阴性对照组、si-LRP16组。将si-LRP16阴性对照序列、si-LRP16分别转染至HCT116细胞,分别命名为阴性对照组和si-LRP16组,未经处理的细胞为空白组。用实时荧光定量聚合酶链式反应(RT-qPCR)测定转染后HCT116细胞LRP16表达水平,四甲基偶氮唑盐微量酶反应比色(MTT)法检测转染后HCT116细胞增殖能力,Transwell小室实验检测转染后HCT116细胞侵袭能力,划痕实验检测转染后HCT116细胞迁移能力,蛋白印迹法(Western blot)测定转染后HCT116细胞中β-catenin表达水平。结果转染后,空白组、阴性对照组和si-LRP16组LRP16表达水平分别为2.17±0.18,2.15±0.19和1.55±0.27,HCT116细胞活性分别为(98.46±3.51)%,(97.92±5.07)%和(71.05±4.73)%;HCT116细胞体外侵袭细胞数分别为(54.90±10.52),(58.79±9.18)和(35.24±8.01)个,HCT116细胞体外迁移能力分别为(90.17±4.08)%,(89.54±6.94)%和(60.18±7.46)%。β-catenin表达水平分别为1.01±0.21,1.01±0.23和0.76±0.14。以上指标,si-LRP16组与空白组和阴性对照组比较,差异均有统计学意义(均P<0.05)。结论抑制LRP16表达可抑制Wnt/β-catenin通路活性,从而降低HCT116细胞增殖、侵袭、迁移能力。

Objective To investigate the effects of human leukemia related protein 16(LRP16) on the proliferation and invasion of colon cancer cells by regulating Wnt/β-catenin pathway. Methods HCT116 cells were assigned to three groups: Blank group,negative control group and si-LRP16 group. The si-LRP16 negative control sequence and si-LRP16 sequence were transfected into HCT116, respectively, named negative control group and si-LRP16 group, and the cells without transfection treatment were used as blank group. The expression of LRP16 were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR). Proliferation of HCT116 were detected by MTT assay. Invasion ability of HCT116 were detected by transwell chamber experiment. Vitro migration capacity of HCT116 were detected by scratch experiments. The expression of β-catenin were detected by Western blot. Results Before transfection,the expressions of LRP16 in blank group,negative control group and si-LRP16 group were 2. 17 ± 0. 18,2. 15 ± 0. 19 and 1. 55 ± 0. 27, respectively;Proliferation of HCT116 were(98. 46 ± 3. 51) %,(97. 92 ± 5. 07) % and(71. 05 ± 4. 73) %, respectively;Invasion ability of HCT11654. 90 ± 10. 52,58. 79 ± 9. 18 and 35. 24 ± 8. 01, respectively;Vitro migration capacity of HCT116 were(90. 17 ± 4. 08) %,(89. 54 ± 6. 94) % and(60. 18 ± 7. 46) %,respectively;The expression of β-catenin were1. 01 ± 0. 21,1. 01 ± 0. 23,0. 76 ± 0. 14,respectively. There were statistically significant differences in the above indices between si-LRP16 group and blank group/negative control group(P < 0. 05). Conclusions Inhibiting the expression of LRP16 could inhibit the activation of Wnt/β-catenin pathway,and thereby decrease the ability of proliferation,invasion and migration in HCT116 cells.