体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

体外不同比例再生肝细胞与结肠癌细胞的共培养

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.     

肝细胞生长因子研究进展

查阅近二十年国外有关HGF/SF文献,基本弄清了HGF/SF的分子结构及组织分布,说明了HGF/SF的调节和cmet受体及两者的关系,揭示了HGF/SF的生物活性、HGF/SF和肿瘤扩散以及与肝再生的关系.因此,HGF/SF在肝再生中起主要作用;是肾再生的关键因子;可促进伤口愈合;通过增加恶性细胞的侵袭性,HGF/SF可能作为肿瘤转移的重要因素之一.     

Co-culture of rat embryos and hepatocytes: in vitro detection of a proteratogen

The technique of whole embryo culture developed by New [Environ Health Perspect 18:105-110, 1976] provides a sensitive assay to evaluate the effects of a test chemical on embryo development independent of maternal influences. To detect proteratogens, this assay must be coupled with an exogenous metabolic activation system. We have developed methods for the co-cultivation of rat embryos with primary hepatocytes, which offers several advantages over subcellular fractions when providing metabolic activation for in vitro assays. In the present study, rat embryos removed from the dam on day 10 of pregnancy were co-cultivated in vitro with primary cultures of rat, rabbit, or hamster hepatocytes. Embryos co-cultivated with hepatocytes developed normally, as did embryos exposed to a test chemical, cyclophosphamide (CP) in the absence of hepatocytes. When embryos were co-cultivated with hepatocytes and exposed to CP, a dose-related embryotoxicity was observed, indicating metabolic activation of the proteratogen. Using hepatocytes isolated from rats pretreated in vivo with phenobarbital, we observed an increase in CP-induced malformations and embryotoxicity compared to those of embryos exposed to CP in the presence of uninduced hepatocytes. The teratogenic bioactivation of CP was inhibited in vitro by the addition of metyrapone. When similar numbers of hepatocytes were used for metabolic activation of CP the induced embryotoxicity was greater in the presence of rabbit and hamster hepatocytes than with rat hepatocytes. Development of procedures for the culture of rat embryos with hepatocytes from other species suggests the utility of this in vitro system for the investigation of species differences in sensitivity to chemical teratogens.