Kail/CD82基因转染对结肠癌细胞株LoVo生物学特性的影响

Changes of biological characteristics of colon carcinoma cell line-LoVo induced by Kai1/CD82 transfection

目的 探讨Kail/CD82对LoVo细胞株生长、黏附、分离与侵袭能力的影响。方法 转染KailcDNA ,建立稳定Kail表达克隆株。采用体外实验的方法 ,绘制转染细胞与对照细胞的生长曲线 ,并检测其黏附、聚集、分离与侵袭能力 ,对比有无差别。结果 与对照细胞相比 ,转染对LoVo细胞的生长无明显影响 ;震荡 10、2 0、3 0、40min时 ,转染细胞与空白对照细胞的黏附细胞数 (× 10 5 )分别为0 0 8、0 63、0 83、0 91和 0 0 4、0 48、0 71、0 82 ,其中震荡 2 0、3 0与 40min时二者差异有显著性 (P <0 0 5) ;震荡 5、10、15min时 ,转染细胞与空白对照细胞的脱落率分别为 13 %、2 0 %、53 %和 11%、2 8%、60 % ,其中震荡 10与 15min时二者的差异有统计学意义 (P <0 0 5) ;5h的温和震荡后 ,转染细胞的聚集体形成率较空白对照细胞为高 (64 8%和 58 6% ,P <0 0 5) ;1d共培养后 ,转染细胞穿过内皮细胞的数目较空白对照细胞的少 (6 3 3 /视野和 17 67/视野 ,P <0 0 5) ;总之 ,转染可以增强细胞的黏附与聚集能力 ,降低其分离与侵袭能力。

Objective To study the influence of Kai1/CD82 transfection on the growth, adherence, separation and invasion potential of LoVo colon carcinoma cell line. Methods Kai1/CD82 cDNA was transfected into LoVo cells, and a stable expressing clone was established. In vitro methodology was used to obtain the growth curve and also to detect the adherence, separation and invasion potential of the transfected LoVo cells, in comparison with those of control cells without transfection. ResultsCompared with the control, no change was observed in the growth pattern of transfected LoVo cells. The numbers of adherent cells in the two groups were 0.08, 0.63, 0.83, 0.91 (×10 5) for the transfected cells and 0.04, 0 48, 0.71, 0.82 (×10 5) for the control cells respectively after 10, 20, 30, 40 minutes culture with shaking. The difference at 20, 30 and 40 minutes was statistically significant (P<0.05). The separation rates of each group were 13%, 20%, 53% for the transfected cells and 11%, 28%, 60% for the control cells, respectively after 5, 10, 15 minutes culture with shaking. The difference at 10 and 15 minutes was statistically significant (P<0.05). The aggregation rate of the transfected cells was higher than that of the control cells after culture with mild shaking for 5 hours (64.8% vs. 58.6%, P<0.05). After co-incuba tion with endothelium cells ECV304, the number of invading cells decreased more in the transfected cells than that in the control cells (6.33/field and 17.67/field,P<0.05). Conclusion Transfection expression of Kail/CD82 into LoVo cell line results in an increase of cell adherence and aggregation, but a diminished capability of separation and invasion, suggesting that the expression of Kai1/CD82 gene may inhibit the metastatic potential of colon carcinoma.