体外与肝细胞共培养对结肠癌细胞移行和增殖的影响

Hepatocytes increase the migration and proliferation of colon cancer cells in vitro

目的探讨肝细胞对结肠癌细胞的移行、增殖能力的影响。方法在体外建立结肠癌细胞与肝细胞三维共培养模型，将结肠癌细胞系Colon-26细胞培养在共培养系统上层，肝细胞培养在下层。实验分为3组：（1）Colon-26细胞与小鼠的肝细胞共培养（肝细胞组）。（2）与成纤维细胞共培养（纤维细胞组）。（3）Colon-26细胞单独培养（对照组）。观察肝细胞对结肠癌细胞的移行、增殖的影响，以及共培养液内表皮细胞生长因子（EGF）、血管内皮生长因子（VEGF）浓度的变化。结果肝细胞组的Colon-26细胞移行能力分别比纤维细胞组和对照组增强（t1=6．296，P=0．000；t2=5．322，P=0．000），Colon-26细胞增殖能力在肝细胞组亦分别比另两组增强（t1=3．905，P=0．005；t2=3．719，P=0．006）。肝细胞组的共培养液中EGF浓度明显升高（z=3．077，P=0．002）。结论在体外能建立结肠癌细胞与肝细胞的三维共培养模型。体外肝细胞能通过分泌生长因子促进结肠癌细胞的移行和增殖。

Objective To investigate the role of hepatocytes in the migration and proliferation of colonic carcinoma cell lines Colon-26 cells by establishing a 3-dimentional coculture system model between colon cancer cells and hepatocytes. Methods Colon-26 cells were cultured in the inserts and hepatocytes were cocultured in the wells. The models were divivided into three groups with group Ⅰ in which mouse hepatocytes were cocuhured with Colon-26 cells; group Ⅱ in which Colon-26 cells were cocultured with fibroblasts ; group Ⅲ in which Colon-26 only were cultured. The migration and proliferation of Colon-26 cells were investigated and the concentration of EGF and VEGF in the cocuhured medium was detected. Results The migration of Colon-26 cells significantly increased in hepatocytes group than in fibroblasts and negative control group respectively（ t1= 6. 296,P = 0. 000; t2 = 5. 322, P = 0. 000）. The proliferation of Colon-26 cells increased in hepatocytes group than the other two groups （ t1 = 3. 905, P = 0. 005 ; t2 = 3. 719, P = 0. 006）. The concentration of EGF increased in hepatocytes group as well （ z = 3.077, P = 0. 002 ）. Conclusions A 3-dimentional cocuhure system model between Colon-26 cells and hepatocytes was established in vitro. Cocultured hepatocytes can promote the migration and proliferation of Colon-26 cells possibly by secreting growth factors into the medium.