VEGFR-3 siRNA腺病毒载体对结肠癌细胞增殖和迁移影响研究

Effect of the adenovirus expressing the short interfering RNA targeting VEGFR-3 on inhibiting the proliferation and migration in colorectal cancer cell

目的探讨血管内皮生长因子受体-3(vascular endothelial growth factor receptor-3,VEGFR-3)siRNA腺病毒载体对结肠癌细胞增殖和迁移的影响。方法将VEGFR-3siRNA腺病毒转染结肠癌LoVo细胞,采用qRT-PCR测定各组结肠癌LoVo细胞VEGFR-3mRNA的表达,蛋白质印迹法检测各组细胞VEGFR-3蛋白的表达,MTT比色法测定各组细胞增殖状况,划痕实验检测LoVo细胞的迁移能力。结果空白对照组VEGFR-3mRNA表达量为0.94±0.12,阴性对照组为0.97±0.14,实验组为0.13±0.19,实验组VEGFR-3 mRNA的表达水平明显降低,F=65.389,P<0.001。蛋白质印迹法检测结果表明,实验组VEGFR-3蛋白的表达量为0.12±0.14,空白对照组为0.94±0.15,阴性对照组为0.97±0.16,实验组VEGFR-3蛋白的表达水平明显下降,F=68.050,P<0.001。空白对照组LoVo细胞增殖活性为0.89±0.03,阴性对照组为0.87±0.05,实验组为0.52±0.06,实验组增殖明显抑制,F=80.135,P<0.001。实验组LoVo细胞相对迁移率为(25.26±1.52)%,空白对照组为(68.24±1.51)%,阴性对照组为(65.18±1.73)%,实验组细胞迁移距离明显缩短,F=63.809,P<0.001。结论腺病毒介导的VEGFR-3siRNA可以通过下调结肠癌细胞VEGFR-3的表达从而抑制结肠癌细胞的增殖和迁移能力。

OBJECTIVE To investigate the effect of the adenovirus expressing the short interfering RNA targeting vascular endothelial growth factor receptor 3 （VEGFR 3） on inhibiting the proliferation and migration in colorectal cancer cell lines LoVo cells. METHODS The adenovirus expressing the small interfering RNA targeting VEGFR-3 was synthe sized and transfected into the LoVo cells. The expressions of VEGFR-3 were measure by qRT-PCR and Western blotting. The proliferation and migration were detected respectively by MTT and Transwell method. RESULTS The expressions of VEGFR 3 mRNA（0.13±0.19） and protein（0. 12±0. 14） were significantly decreased by the recombinant adenovirus （F=65. 389,P〈0. 001;F=68. 050,P〈0. 001） ,compared with the naked control group （0.94±0.12; 0. 94±0.15） and negative control group（0.97±0.14; 0.97 ± 0.16）. The proliferation was significantly decreased by the recombinant adenovires（0.52 ±0.06） （F= 80. 135, P〈0.001）, compared with the naked control group（0.89 ± 0.03） and negative control group（0.87± 0.05）. The migration was significantly decreased by the recombinant adenovirus （25. 26 ± 1. 52）% （F= 63. 809,P〈 0. 001 ） , compared with the naked control group （68. 24 ± 1. 51） % and negative control group （ 65. 18 ± 1.73） %, CONCLUSION The adenovirus expressing the short interfering RNA targeting VEGFR-3 could inhibit the proliferation and migration in colorectal cancer cells.