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.     

全反式维甲酸对人结肠癌LoVo细胞化疗药物敏感性及Survivin基因表达的影响

Objective To study the effects of all-trans-retinoic acid (ATRA) on chemotherapeutics sensibility and expression of Survivin in human LoVo colon cancer cell line, and to provide a theoretical basis for clinical application of ATRA.Methods Flow cytometer (FCM) was used to detect the cell cycle of LoVo colon cancer cell line after treated with various dose of ATRA (10-5 mol/L, 10-6 mol/L and 10-7 mol/L)for 24 h, 48 h, 72 h, 5 d, 9 d and 15 d respectively.The cells were divided into ATRA group (10-6 mol/L)and control group (cultured in routine medium RPMI1640).Meanwhile, each group were divided into subgroups respectively treated with 0 g/L, 2 g/L, 4 g/L and 6 g/L 5-fluorouracil(5-FU) , or with 0 mg/L, 200 mg/L, 400 mg/L and 600 mg/L mitomycin (MMC).MTT assay was used to detect the viability of cells so as to analyze the change in chemotherapeutics sensibility.Drug interactions were analyzed according to the interaction effects.Staining with acridine orange (AO) and ethidium bromide (EB) was used to study the apoptosis in control group(RPMI1640) , ATRA group (10-6 mol/L), 5-FU group (4 g/L), MMC group (400 mg/L), ATRA (10-6 mol/L) + 5-FU (4 g/L) group and ATRA (10-6 mol/L) + MMC (400 mg/L) group.The expression of Survivin in the LoVo cell line was investigated by immunofluorescence technique.Results Compared with the control group, the percentage of cell in Go-G1 phase increased after interference with various doses of ATRA for 24 hours, peaked at 48 h, and gradually decreased afterwards, while the percentage of cells in stages S and G2-M decreased, reach the trough at 48 h, and then increased gradually.Effects of 10-6 mol/L ATRA were most significant at all time spots.Compared with control group, 10-6 mol/L ATRA reduced cell survival rate (P＜0.05),and the survival rates in control subgroups treated with various doses of 5-FU were significantly higher than those in ATRA group (all P＜0.05).In MMC-treated control group, cell survival rate with 200 mg/L MMC was lower than that in ATRA group (0.72±0.02 vs 1.03±0.13, P＜0.05) , while treatment with 400 mg/L or 600 mg/L MMC did not result in lower cell survival compared with the ATRA group (0.52±0.05 vs 0.39±0.02, 0.36±0.02 vs 0.36±0.01, all P>0.05).Positive interactions of 10-6 mol/L ATRA with 5-FU (2 g/L, 4 g/L, 6 g/L) or MMC (200 mg/L, 400 mg/L) were found.Under fluorescence microscope, cells in ATRA + 5-FU group and ATRA + MMC group showed significantly small size, pyknosis and chromatin condensation and formation of apoptotic bodies when compared with control group, ATRA group, 5-FU group and MMC group.Expression of Survivin was inhibited after 10-6 mol/L ATRA treatment for 48 h compared with control group (33.33% vs 27.96% , P＜0.05).Conclusion ATRA enhances the chemosensibility of LoVo colon cancer cell line to 5-FU and MMC,and induces cell apoptosis, potentially through inhibition of Survivin gene expression.     

Effect of He-Ne Laser on the Expressions of EGF and EGFR in Gastric Mucosa of Chronic Atrophic Gastritis

To investigate the histological change and the expressions of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) in atrophic gastric mucosa in rats so as to appraise the effect of these regulators on the formation of atrophic gastritis, to study the expressions of EGF and EGFR in rats with chronic atrophic gastritis (CAG) after the irradiation of He-Ne laser, and to investigate the relations of He-Ne laser and precancerous lesion or apoptosis. The rats were divided into normal group, model group and laser group. The models of CAG rats were established with enema which was the mixed liquor consisted of sodium salicylate and alcohol, combined with irregular fasting and compulsive sporting as pathogenic factors. He-Ne laser(3.36 J/cm2 ) was used to irradiate CAG rats, once a day for 7 min, 20 days as a course of treatment; the expressions of EGF and EGFR were detected with immunohistochemical method. During the process of antral atrophy, the expressions of EGF and EGFR increased, they were higher in model group than those in normal group (P<0.05). After the irradiation of He-Ne laser, the expressions of EGF and EGFR were obviously lower than those in model group (P<0.05). We draw the conclusion as follows: gastric mucosa of model rat is in a hyper-proliferation status, with high protein expressions of apoptosis suppressor EGF and EGFR. CAG has some correlation with the imbalance between cell proliferation and apoptosis. He-Ne laser (3.36 J/cm2 ) can reduce the expressions of EGF and EGFR, which is an effective physiostimulator to stimulate the gastric mucosa of rat. The appropriate secretions of EGF and EGFR are propitious to the repair and regeneration for the gastric mucosa tissues, thus preventing CAG to canceration.