CAJ | 학술논문

目的观察体外与不同比例再生肝细胞共培养对结肠癌细胞增殖潜能及肝再生相关因子的影响.方法 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,分别以10:1(A组)、1:1(B组)与人结肠癌细胞株SW480共培养,对照组为单独培养的结肠癌细胞(C组).培养后第24和72 h,通过3H-胸腺嘧啶核苷(3H-TdR)掺人法比较各组培养后的增殖能力,并采用酶联免疫吸附试验(ELISA)检测表皮生长因子(EGF)、胰岛素样生长因子-1(IGF-1)及肝细胞生长因子(HGF)的浓度.结果培养后24 h 3组间3H-TdR掺人率和EGF、IGF-1、HGF的浓度差异无统计学意义(P>0.05),72 h A、B两组的3H-TdR掺入率(15.9±1.4,13.2±1.5)和EGF[(722.9±55.4)ng/L,(498.2±41.5)ng/L]、IGF-1[(755.2±35.7)ng/L,(538.1±37.5)ng/L]明显高于C组(P<0.05),且A组高于B组(P<0.05).HGF的浓度在培养后第24、72小时差异均无统计学意义(P>0.05).结论与再生肝细胞共培养的结肠癌细胞增殖能力增强,其机制可能与来自于肝细胞内的生长信号增加结肠癌细胞EGF和IGF-1的表达有关.
목적 관찰체외여불동비례재생간세포공배양대결장암세포증식잠능급간재생상관인자적영향.방법 70%간절제대서모형술후24 h채용원위효원매관류법분리획득유활성적재생간세포,분별이10:1(A조)、1:1(B조)여인결장암세포주SW480공배양,대조조위단독배양적결장암세포(C조).배양후제24화72 h,통과3H-흉선밀정핵감(3H-TdR)참인법비교각조배양후적증식능력,병채용매련면역흡부시험(ELISA)검측표피생장인자(EGF)、이도소양생장인자-1(IGF-1)급간세포생장인자(HGF)적농도.결과 배양후24 h 3조간3H-TdR참인솔화EGF、IGF-1、HGF적농도차이무통계학의의(P>0.05),72 h A、B량조적3H-TdR참입솔(15.9±1.4,13.2±1.5)화EGF[(722.9±55.4)ng/L,(498.2±41.5)ng/L]、IGF-1[(755.2±35.7)ng/L,(538.1±37.5)ng/L]명현고우C조(P<0.05),차A조고우B조(P<0.05).HGF적농도재배양후제24、72소시차이균무통계학의의(P>0.05).결론 여재생간세포공배양적결장암세포증식능력증강,기궤제가능여래자우간세포내적생장신호증가결장암세포EGF화IGF-1적표체유관.
Objective To investigate the proliferation potential of colon cancer cells co-cultured with different proportions regenerating hepatocytes and role of the cell growth factors related to liver regeneration in vitro.Methods Active regenerating hepatocytes were obtained by collagenase perfusion in situ of rats models underwent 70% liver resection after 24 hours.Co-cultures with different proportions of hepatocytes to human colon cell line SW480 were performed respectively.The proportion of hepatocytes to human colon cell line SW480 Was 10:1 in group A,1:1 in group B.There was only human colon cell line SW480 in group C(control group).Proliferation capacity was assessed with the percentage by 3H-thy incorporation.Concentration of epidermal growth factor(EGF),insulin-like growth factor-1(IGF-1)and hepatocyte growth factor(HGF)was analyzed by ELISA.Results There was no significant difference among the 3 groups in 3H-thy incorporation percentage,and concentration of EGF,IGF-land HGF after 24 hours' culture(P>0.05).3H-thy incorporation percentage(15.9±1.4,13.2±1.5),and concerntration of EGF [(722.9±55.4)ng/L,(498.2+41.5)ng/L]and IGF-1[(755.2±35.7)ng/L,(538.1±37.5)ng/L]in Group A and B were higher than that in group C after 72 hours(P<0.05).Especially the indexes above in group A were higher than that in group B(P<0.05).there was no significant difference in concerntration of HGF among the 3 groups after 72 hours(P>O.05).Conclusions These results imply that the regenerating hepatocytes contribute to the hyperproliferative state of co-culturing colon cancer cells.The mechanism maybe have relationship with EGF and IGF-1 high expression in colon cancer cells caused by growth signals coming from hepatocytes.