CAJ | 학술논문

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.
목적 통과계제전이법재라서중사선출인결장암간전이세포,연후대결장암간전이세포적생장증식능력진행감정.방법 통과SW1116세포주라서피하련속5차전대후취피하충식류라서결장원위충식,분별취간장전이조화피하5대세포진행원대배양,획득대장암간전이세포화피하5대적원대세포,분별명명위결직장암간전이세포1대(CHM-1)화피하제5대(P5),통과CCK-8측생장곡선、류식검측세포주기이급세포주기상관적단백적면역세포화학래대SW1116、CHM-1화P5증식능력진행검측.결과 세포증식실험현시CHM-1생장속도명현만우SW1116화P5세포(P＜0.05);류식세포주기분석중CHM-1세포G41기(68.390±2.865)%비례고우SW1116(50.440±1.472)%화P5(53.930±2.651)%(P＜0.05),G2기CHM-19(13.530±4.411)%비례요저우SW1116(32.030±5.645)%화P5(29.720±3.559)%(P＜0.05),S기3충세포차이무통계학의의(18.250±6.901)%、(19.050±4.162)%、(18.930±0.169)%(P＞0.05);면역세포화학분석발현세포주기단백Cyclin DI재CHM-1(23.7±4.7)%중적표체비SW1116(30.2±5.1)%화P5(32.1±4.2)%감소(P＜0.05).결론 발현침습능력화생장증식능력구유반향관련,즉결장암간전이세포구유경강적침습능력,단생장증식속도비기타세포경만.
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.