| Objective: The Lewis lung cancer mice act as experimental animal models,detecting the expression charactistic of Fas,Fas L,Caspase-8,Caspase-3,Bcl-2 and Bax in Lewis lung cancer tissues by immunohistochemistry and Western blotting.Meanwhile exploring the anticancer mechanism of “lung cancer Ⅰ prescription”,and provide a reliable scientific foundation for its clinical application.Method: A total of 48 Lewis lung cancer mice models were selected and randomly divided into four groups,respectively the model group,lung cancer I prescription group,Cisplatin group and combination group(Cisplatin+lung cancer Ⅰ prescripttion).the drug was given mice by intragastric administration of normal saline once a day and intraperitoneal injection of normal saline once time per three days in the model group.Those mice were intragastrically administrated with lung cancer Ⅰ prescription once a day and intraperitoneal injection of normal saline once time per three days in the lung cancer Ⅰ prescription group.Those mice was given by intragastric administration of normal saline once a day and intraperitoneal injection of Cisplatin once time per three days in the Cisplatin group.Those mice was given lung cancer Ⅰ prescription once a day by intragastric administration and intraperitoneal injection of Cisplatin once time per three days in the combination group.During the period of experiment,the physiological state of mice was observed.12 days later,they were sacrificed by using cervical dislocation.The tumor tissue was taken out and weighed,then the tumor inhibition rate was calculated.Histopathological slices were performed by HE staining,and protein expressions of Fas,Fas L,caspase-8,caspase-3,bcl-2 and Bax were detected by immunohistochemistry and western blotting.The experimental data are expressed as mean ± standard deviation,using SPSS 20.0 statistical software,the experimental data are selected as one-way ANOVA.Result: 1.The quality of life:Compared with the model group,the activity and diet of the mice in each drug group improved.Compared with the three experimental groups,the survival status of mice in the combination group was significantly better than them in the lung cancer I prescription group and the Cisplatin group.The survival status of mice in the Cisplatin group was worse than them in the lung cancer I prescription group.2.Results of tumor weight and tumor inhibition rate: 2.1 Tumor weight: Compared with the model group,the mean tumor weight in each drug group decreased,and the difference was statistically significant(P<0.05).Compared with the three experimental groups,the mean tumor weight in the cisplatin group was lower than than it in the lung cancer I prescription group,the difference was statistically significant(P<0.05);the mean tumor weight in the combination group was lower than than it in the Cisplatin group,the difference was statistically significant(P<0.05).2.2 Tumor inhibition rate:The tumor inhibition rate of combination group was 40.1%,which was the best;the tumor inhibition rate of Cisplatin group was second,28.5%;the tumor inhibition rate of lung cancer I prescription group was the lowest,15.2%.3.Histopathological observations: Compared with the model group,the density of tumor cells in other three groups decreased,cell edema was significant,necrosis area increased,and necrosis structure was mostly flaky.In the combination group have the most tumor cell necrosis area and cell edema.And Compared with lung cancer I prescription group,the cisplatin group had larger necrosis area,wider necrosis area and more significant degree of cell edema.4.The results of immunohistochemistry showed that compared with the model group,the proteins expression of Fas,Fas L,Caspase-8,Caspase-3 and Bax was increased and the proteins expression of Bcl-2 was decreased in tumor tissues of three drug groups,the difference was statistically significant(P<0.05);Compared with the lung cancer I prescription group,the proteins expression of Fas L,Caspase-3 and Bax in the tumor tissues of the cisplatin group and the combination group was increased,and the proteins expression of Bcl-2 was decreased,the difference was statistically significant(P<0.05).The proteins expression of Fas and Caspase-3 in tumor tissues of cisplatin group was higher than them in lung cancer I prescription group,but the difference was not statistically significant(P>0.05).Compared with the lung cancer I prescription group,the proteins expression of Fas and Caspase-8 in the tumor tissue of the combination group was increased,the difference was statistically significant(P<0.05).Compared with the cisplatin group,the proteins expression of Fas,Fas L,Caspase-8,Caspase-3 and Bax in the tumor tissue of the combination group was increased,and the proteins expression of Bcl-2 was decreased,the difference was statistically significant(P<0.05).5.Compared with the model group,Western blotting showed that the proteins expression of Fas,Fas L,Caspase-8,Caspase-3 and Bax increased and the proteins expression of Bcl-2 decreased in the tumor tissues of other three groups,the difference was statistically significant(P<0.05).Compared with the lung cancer I prescription group,the proteins expression of Fas L,Caspase-8,Caspase-3 and Bax in the tumor tissues of the cisplatin group and the combination group was increased,the difference was statistically significant(P<0.05).The proteins expression of Fas in the cisplatin group was higher than them in the lung cancer I prescription group,but the difference was not statistically significant(P>0.05).The proteins expression of Fas in the tumor tissue of the combination group was higher than it in the lung cancer I prescription group,and the difference was statistically significant(P<0.05).The proteins expression of Bcl-2 in the tumor tissue of the combination group was lower than it in the lung cancer I prescription group,and the difference was statistically significant(P<0.05).There was not statistically significant for the proteins expression of Bcl-2 in the tumor tissue of cisplatin group make a comparison of lung cancer I prescription group,(P>0.05);Compared with the cisplatin group,the proteins expression of Fas,Fas L,Caspase-8,Caspase-3 and Bax in the tumor tissue of the combination group were increased,and the proteins expression of Bcl-2 was decreased,the difference was statistically significant(P<0.05).Conclusion: 1.The Lung I cancer prescription can increase the proteins expression of Fas,Fas L and Caspase-8(the apoptosis pathway of death receptors)in tumor tissues,it suggests that its anti-cancer mechanism is to promote apoptosis of lung cancer cells.2.The Lung cancer I prescription can increase the proteins expression of Bax,Caspase-3 and decrease proteins the expression of Bcl-2(the mitochondrial apoptotic pathway)in tumor tissues,it suggests that its anti-cancer mechanism is also to promote apoptosis of lung cancer cells.3.In tumor tissues,the proteins expression of Fas,Fas L,Caspase-8,Bax and Caspase-3 in the combination group was increased,and the proteins expression of Bcl-2 was decreased,it suggests that Lung cancer I prescription combined with cisplatin can enhance the efficacy.Combined with the results of previous experiments,lung cancer I prescription combined with Cisplatin has a strong anti-tumor effect.However,in clinical practice,its safety in combination with chemotherapeutic drugs or targeted drugs is not clear.It is necessary to go on the study of toxic and side effects of chemotherapeutic drugs and targeted drugs,which has certain practical significance for further promotion in the clinical. |
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