肾母细胞瘤过表达基因对肾癌细胞的作用

The role of nephroblastoma overexpressed gene on renal cell carcinoma

目的探讨肾母细胞瘤过表达（nephroblastomaoverexpressed，NOV）基因对肾癌细胞增殖、黏附、侵袭和迁移能力的影响。方法构建NOV蛋白表达真核细胞重组表达质粒pEGFP—C1-NOV，转染人肾癌细胞株786—0。通过计数细胞绘制生长曲线、水溶性四氮唑法（WST-1）检测细胞生长抑制率，通过细胞黏附实验、侵袭实验和细胞迁移实验，比较转染pEGFP—C1-NOV组（实验组）与转染空载体组（空载组）及未转染组（空白组）的细胞增殖、黏附、侵袭和迁移能力的差异。结果与空白组比较，实验组48、72h抑制率分别为29．14％、32．46％，空载组分别为9．25％和-8．16％，实验组与空白组比较差异有统计学意义（P〈0．05），空载组与空白组相比差异无统计学意义（P〉0．05）。与层粘连蛋白黏附，实验组A值为0．26±0．03，高于空白组（0．15±0．01）和空载组（0．14±0．02），差异有统计学意义（P〈0．05）；与人纤维连接蛋白黏附，实验组A值为0．28±0．04，高于空白组（0．124±0．095）和空载组（0．128±0．082），差异有统计学意义（P〈0．05）。实验组穿越Matrigel基质胶的细胞数为240．25±23．12，显著高于空白组（56．16±6．25）和空载组（50．28±7．13），差异有统计学意义（P〈0．05）；实验组迁移通过聚碳酸酯微孔滤膜的细胞数为267．25±20．94，显著高于空白组（66．10±5．68）和空载组（56．28±4．11），差异有统计学意义（P〈0．05）。结论NOV可抑制。肾癌细胞的增殖，促进。肾癌细胞786—0的黏附、侵袭和迁移。

Objective To investigate the effects of nephroblastoma overexpressed （ NOV ） on proliferation, adhesion, migration and invasion of remal cell curcinomai （RCC） cells. Methods We constructed a NOV expression plasmid and transfeeted the plasmid into RCC cell line 786-0 and analyzed the effects of NOV expression on proliferation, adhesion, migration and invasion of RCC cells by growth curve assay, WST-1 assay, cell adhesion assay, matrigel invasion assay and transwell migration assay. Results The stable NOV transfected 786-0 cells （786-O-NOV） showed decreased growth rate, at 48 h and 72 h, the proliferation activities of 786-O-NOV cells were inhibited by 29.14% and 32.46% the proliferation activities of empty vector cells were inhibited by 9.25% and -8. 16%, respectively, compared to 786-0 cells （P 〈 0. 05） ; while the 786-0 cells transfected with empty vector （786-O-mock） had no difference with 786-0 cells. Adhesion assay indicated significantly increased adhesion of 786-O-NOV cells to fihronectin （0.26 ＋ 0.03） and laminin （0.28 ±0.04）, compared to 786-0 cells （0.15 ±0.01, 0.12±0.10） and 786-0- mock cells （0.14 v 0.02, 0. 13 v 0.08）. Invasion assay displayed that the numbers of cells penetrated through matrigel membrane were significantly higher in 786-O-NOV cells （240.25 ± 23.12） compared to 786-0 cells （56. 16 ± 6.25） and 786-O-mock cells （50.28 ± 7. 13）. Migration assay displayed that the numbers of cells passed through polycarbonate filters were significantly higher in 786-O-NOV cells （267.25v 20.94） compared to 786-0 cells （66. 10 ±5.68 ） and 786-O-mock cells （ 56.28 ± 4.11 ）.Conclusion NOV exhibits antiroliferative effects on RCC cells; however, it promotes adhesion, migration and invasion of RCC cells.