CAJ | 학술논문

目的探讨与再生肝细胞共培养时结肠癌细胞增生潜能的改变及其可能机制.方法人结肠痛细胞株SW480,与70%肝切除大鼠模型术后24 h采用原位胶原酶灌流法分离获得有活性的再生肝细胞以不同比例混合进行体外培养;采用[3H].胸腺嘧啶核苷(3H-TdR)掺入率和Westernblot检测细胞培养24、72、96和120 h的增生反应及表皮生长因子受体(EGFR)、胰岛素样生长因子-1受体(IGF-1R)和肝细胞生长因子受体(c-met)表达变化.结果 1:1和1:10比例共培养组结肠癌细胞增生反应增强,3H-TdR掺人率从培养第72 h开始增加,至第120 h呈持续增加趋势(P<0.05),EGFR和IGF-1R从第24 h开始表达增强,且表达水平呈时间效应关系,c-met表达无明显变化;10:1比例共培养组结肠癌细胞增生情况、3种受体表达水平均无明显变化.结论与再生肝细胞共培养可增强结肠癌细胞EGFR、IGF-1R的表达,方式可能来自于肝细胞内的生长信号通过旁分泌机制增强结肠癌细胞EGFR和IGF-1R的表达,从而促进结肠癌细胞的增生反应.
목적 탐토여재생간세포공배양시결장암세포증생잠능적개변급기가능궤제.방법 인결장통세포주SW480,여70%간절제대서모형술후24 h채용원위효원매관류법분리획득유활성적재생간세포이불동비례혼합진행체외배양;채용[3H].흉선밀정핵감(3H-TdR)참입솔화Westernblot검측세포배양24、72、96화120 h적증생반응급표피생장인자수체(EGFR)、이도소양생장인자-1수체(IGF-1R)화간세포생장인자수체(c-met)표체변화.결과 1:1화1:10비례공배양조결장암세포증생반응증강,3H-TdR참인솔종배양제72 h개시증가,지제120 h정지속증가추세(P<0.05),EGFR화IGF-1R종제24 h개시표체증강,차표체수평정시간효응관계,c-met표체무명현변화;10:1비례공배양조결장암세포증생정황、3충수체표체수평균무명현변화.결론 여재생간세포공배양가증강결장암세포EGFR、IGF-1R적표체,방식가능래자우간세포내적생장신호통과방분비궤제증강결장암세포EGFR화IGF-1R적표체,종이촉진결장암세포적증생반응.
Objective To investigate the stimulated proliferation of colon cancer cells in co-cultures of regenerating hepatocytes. Methods Regenerating hepatocytes(24 hours after partial hepatectomy)were obtained by collagenase perfusion of models of rats undergoing 70% liver resection. To determine whether the ratio of human colon cell line SW480 cells to hepatocytes in co-cultures has influence on their interaction,these cells were cultured in ratios of 1: 101:1, or 10: 1. Proliferation capacity was assessed by the percentage of 3 H-TdR incorporation. Expression of epidermal growth factor receptor(EGFR), insulin-like growth factor1 receptor(IGF-1R)and hepatocyte growth factor receptor(c-met) were analyzed by western blot. Results For co-cultured SW480 and hepatocytes in the ratios of 1: 1 and 1: 10, an increase of disintegrations per minute(dpm) occurred after 72 hours' culture, and lasted at 120 hours' culture(P < 0.05). No difference was found between the group with ratio of 10:1 and control group. Protein levels of EGFR and IGF-1R, but not c-met, were significantly increased between culture of 24 hours and 120 hours; however, no change of these receptors was found in the ratio of 10: 1. Conclusions These results imply that co-culturing human colon cancer cells with regenerating hepatocytes leads to increased expression of EGFR and IGF-1R. We conclude that this effect is probably dependent on paracrine stimulation, by which numerous signals from the hepatocytes contribute to the hyperproliferative state of colon cancer cells via up-regulating the responding receptors.