| Objective:To observe the efficacy and safety of via micro pump and intraperitoneal injection of recombinant human endostatin(endostar)combined with cisplatin on Lewis lung cancer xenografts in mice.Methods:Establishment of a model of Lewis lung cancer xenografts in C57BL/6J mice,the mice were divided into six groups randomly:(1)saline:daily intraperitoneal injection of 0.2mL saline for 14 consecutive days;(2)saline plus intraperitoneal implantation osmotic pump:daily intraperitoneal injection of 0.2mL saline for 14 consecutive days.Abdominal implantation of ALZET osmotic pumps in mice for 14 days,pumps were replaced every 7 days;(3)endostar[5mg/(kg·d),ip]plus cisplatin:intraperitoneal injection of mice 5mg/(kg d)for 14 consecutive days,and then mice were intraperitoneally injected with cisplatin 3mg/(kg d)for 3 days;(4)endostar[10mg/(kg·d),ip]plus cisplatin:intraperitoneal injection of mice 10mg/(kg·d)for 14 consecutive days,and then mice were intraperitoneally injected with cisplatin 3mg/(kg·d)for 3 days;(5)endostar[20mg/(kg d),ip]plus cisplatin:intraperitoneal injection of mice 20mg/(kg d)for 14 consecutive days,and then mice were intraperitoneally injected with cisplatin 3mg/(kg d)for 3 days;(6)endostar[5mg/(kg·d),pump]plus cisplatin:mice were treated 5mg/kg of endostar daily by mini-osmotic pumps for 14 days,and then were given 3mg/kg of cisplatin by intraperitoneal injection for 3 days.After drug administration,the signs and behaviors of the mice were observed dynamically,the volume of tumor in different groups was measured.Determination of the microvascular density(MVD)in tumor tissues by immunohistochemistry,and the serum VEGF levels were evaluated by ELISA.Blood samples of mice were collected for blood routine and biochemical examination,and the stability of endostar in the osmotic pump was detected.Results:1.The establishment of tumor growth curves:Tumor volume measurements showed that the difference in tumor volume between the groups was statistically significant at the end of the experiment(F=1803.161,P<0.001).The growth rate of the tumor group was faster in saline group and saline plus intraperitoneal implantation osmotic pump group.There was no significant difference in tumor volume at the end of the experiment(t=-2.288,P=0.391).The tumor growth of each endostar plus cisplatin group was slow.The tumor volume at the end of the experiment was smaller than that of saline group,and the difference was statistically significant(all P<0.001).At the end of the experiment,the tumor volume of endostar[5mg/(kg d),pump]plus cisplatin group was less than the endostar[5mg/(kg·d),ip]plus cisplatin group(r=-31.1 50,P<0.001)and endostar[10mg/(kg d),ip]plus cisplatin group(t=-17.730,P<0.001),there was no significant difference in tumor volume compared with endostar[20mg/(kg d),ip]plus cisplatin group(t=-1.948,P=0.850).2.The stability of endostar in the osmotic pump:Endostar samples were analyzed for proteolytic degradation by SDS-PAGE under reducing conditions.Wells were loaded with equal concentrations of endostar sample.Notice that no degradation is apparent compared with the sample stored at 4?.3.The microvascular density in tumor tissues in different groups:Immunohistochemistry results showed that there was a statistically significant difference in microvascular density between the tumor groups(F=176.588,P<0.001).There was no significant difference between saline group and saline plus intraperitoneal implantation osmotic pump group(t=2.249,P=0.429).Compared with saline group,the MVD of each endostar plus cisplatin group decreased(all P<0.001).The MVD counts of endostar[5mg/(kg d),pump]plus cisplatin group were lower than endostar[5mg/(kg·d),ip]plus cisplatin group(t=-14.460,P<0.001)and endostar[10mg/(kg d),ip]plus cisplatin group(t=-8.813,P<0.001),there was no significant difference compared with endostar[20mg/(kg·d),ip]plus cisplatin group(t=-1.882,P=0.979).4.The serum VEGF levels in different groups:The results of the ELISA test showed that there were significant differences in serum VEGF expression levels between the groups(F=595.805,P<0.001).There was no significant difference between saline group and saline plus intraperitoneal implantation osmotic pump group(t=1.479,P>0.999).Compared with saline group,the expression of VEGF in each endostar plus cisplatin group was decreased(all P<0.001).The expression of VEGF in endostar[5mg/(kg·d),pump]plus cisplatin group was lower than that of endostar[5mg/(kg·d),ip]plus cisplatin group(t=-14.330,P<0.001)and endostar[10mg/(kg·d),ip]plus cisplatin group(t=-8.132,P<0.001),there was no significant difference compared with endostar[20mg/(kg·d),ip]plus cisplatin group(t=-0.626,P>0.999)5.Toxicity observation:In saline group and saline plus intraperitoneal implantation osmotic pump,the mice were lively and energetic in the early stage,and the spirit was slightly poor,the fur was bright,and the diet was normal.During the endostar plus cisplatin groups,the mice had poor spirits,dull skin and reduced diet,there were 4 to 5 mice with beards in each group,which may be related to chemotherapy drugs.Related blood index testing showed that,there were 6 mice in endostar[5mg/(kg·d),ip]plus cisplatin group with decreased white blood cell and/or platelet count,and there were 4 mice in endostar[5mg/(kg·d),pump]plus cisplatin group,there was no significant difference between the two groups(P=0.656).The levels of AST and/or ALT were increased in both groups,but the difference was not statistically significant(P>0.999).Conclusion:1.Endostar anti-angiogenesis combined with cisplatin can effectively inhibit tumor growth.The effect of endostar via micro pump is better than intraperitoneal injection on Lewis lung cancer xenografts in mice2.Endostar remained stable and active in osmotic pumps for at least 7 days.Notice that no degradation is apparent compared with the sample stored at 4?.3.Endostar via micro pump can significantly inhibit tumor tissue microvessel formation compared with intraperitoneal injection and promote decreased serum VEGF expression.4.Endostar via micro pump can reduce the dosages of drug without increasing drug toxicity. |
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