结肠癌肝转移细胞增殖能力减弱

目的 通过阶梯转移法在裸鼠中筛选出人结肠癌肝转移细胞,然后对结肠癌肝转移细胞的生长增殖能力进行鉴定.方法 通过SW1116细胞株裸鼠皮下连续5次传代后取皮下种植瘤裸鼠结肠原位种植,分别取肝脏转移灶和皮下5代细胞进行原代培养,获得大肠癌肝转移细胞和皮下5代的原代细胞,分别命名为结直肠癌肝转移细胞1代(CHM-1)和皮下第5代(P5),通过CCK-8测生长曲线、流式检测细胞周期以及细胞周期相关的蛋白的免疫细胞化学来对SW1116、CHM-1和P5增殖能力进行检测.结果 细胞增殖实验显示CHM-1生长速度明显慢于SW1116和P5细胞(P＜0.05);流式细胞周期分析中CHM-1细胞G41期(68.390±2.865)%比例高于SW1116(50.440±1.472)%和P5(53.930±2.651)%(P＜0.05),G2期CHM-19(13.530±4.411)%比例要低于SW1116(32.030±5.645)%和P5(29.720±3.559)%(P＜0.05),S期3种细胞差异无统计学意义(18.250±6.901)%、(19.050±4.162)%、(18.930±0.169)%(P＞0.05);免疫细胞化学分析发现细胞周期蛋白Cyclin DI在CHM-1(23.7±4.7)%中的表达比SW1116(30.2±5.1)%和P5(32.1±4.2)%减少(P＜0.05).结论 发现侵袭能力和生长增殖能力具有反向关联,即结肠癌肝转移细胞具有更强的侵袭能力,但生长增殖速度比其他细胞更慢.

Abstract：

Objective To study the proliferation ability of liver metastatic colorectal cancer cells by establishing stepwise metastatic colorectal carcinoma cell model via in vivo selection.Methods SW1116 cells were transplanted into the subcutis of nude mice and serially passaged for 5 times,and then the subcutaneous tumor was implanted into the cecal wall of new nude mice.The liver metastatic lesions and the subcutaneous carcinoma cells of the fifth generation were picked for primary culture.Two cell lines named CHM-1 and P5 were obtained.Proliferation levels were analyzed by testing of cell growth curve,cell cycle analyses by flow cytometry and Cyclin D1 analysis by immunocytochemistry.Results CHM-1 grew much slower than P5 and SW 1116 although there was no difference between P5 and SW 1116 in cell proliferation assay ( P＜0.05 ).Cell cycle analysis revealed that the percentage of cells in G0/G1 phase was higher (P＜0.05) in CHM-1 (68.390 ±2.865)% than in SW1116 (50.440 ± 1.472)% and P5 (53.930 ± 2.651)%,but that in G2/M phase was lower ( P＜0.05 ) in CHM-1 ( 13.530 ± 4.411 )% than in SW1116 (32.030 ±5.645)% and P5 (29.720 ±3.559)% rough there was no difference in S phase ( 18.250 ± 6.901 )%,( 19.050 ± 4.162 )%,( 18.930 ± 0.169 )% ( P ＞ 0.05 ).The expression of cell cycle proteins Cyclin D1 was decreased ( P＜0.05 ) in CHM-1 ( 23.7 ± 4.7 )% as compared with thoseinP5(32.1±4.2)% and SW1116 (30.2 ±5.1)%.Conclusion In the same cell line,there was a reverse relationship between the ability of invasion and proliferation.

Alleviated proliferation in liver metastatic colorectal cancer cells

Objective To study the proliferation ability of liver metastatic colorectal cancer cells by establishing stepwise metastatic colorectal carcinoma cell model via in vivo selection.Methods SW1116 cells were transplanted into the subcutis of nude mice and serially passaged for 5 times,and then the subcutaneous tumor was implanted into the cecal wall of new nude mice.The liver metastatic lesions and the subcutaneous carcinoma cells of the fifth generation were picked for primary culture.Two cell lines named CHM-1 and P5 were obtained.Proliferation levels were analyzed by testing of cell growth curve,cell cycle analyses by flow cytometry and Cyclin D1 analysis by immunocytochemistry.Results CHM-1 grew much slower than P5 and SW 1116 although there was no difference between P5 and SW 1116 in cell proliferation assay ( P＜0.05 ).Cell cycle analysis revealed that the percentage of cells in G0/G1 phase was higher (P＜0.05) in CHM-1 (68.390 ±2.865)% than in SW1116 (50.440 ± 1.472)% and P5 (53.930 ± 2.651)%,but that in G2/M phase was lower ( P＜0.05 ) in CHM-1 ( 13.530 ± 4.411 )% than in SW1116 (32.030 ±5.645)% and P5 (29.720 ±3.559)% rough there was no difference in S phase ( 18.250 ± 6.901 )%,( 19.050 ± 4.162 )%,( 18.930 ± 0.169 )% ( P ＞ 0.05 ).The expression of cell cycle proteins Cyclin D1 was decreased ( P＜0.05 ) in CHM-1 ( 23.7 ± 4.7 )% as compared with thoseinP5(32.1±4.2)% and SW1116 (30.2 ±5.1)%.Conclusion In the same cell line,there was a reverse relationship between the ability of invasion and proliferation.