Effect And Mechanism Of Jinlong Capsule On The Proliferation And Apoptosis Of MGC-803 And BGC-823 Cells In Gastric Cancer In Vivo And In Vitro

Cancer is one of the most lethal diseases in the world.Gastric cancer is one of the most common digestive system tumors in malignant tumors.Its morbidity and mortality are still high.Most of the patients died of gastric cancer were in advanced stage,and lost the chance of surgery.The prognosis of non-surgical treatment was also bad.Apoptosis is also called Programmed Cell Death(PCD).Apoptosis is one of the basic characteristics of life,it can maintain the normal growth and development of the body and the stability of the internal environment together with cell proliferation.The uncontrolled proliferation and apoptosis of cells will lead to many diseases.The discovery of caspases,a family of cysteine proteases,makes people realize that apoptosis is caused by the activation of caspases,which leads to the activation of proteolytic system.Caspases is the core of apoptosis.Activating or inhibiting caspases system can induce or inhibit apoptosis,which becomes a new way to treat apoptosis-related diseases.Survivin,Bcl-2-associated X protein(Bax)and B-cell lymphoma-2(Bcl-2)also play important roles in cell apoptosis.In recent years,the search for anti-tumor drugs from Chinese herbal medicine has attracted extensive attention.Jinlong Capsule(JLC),a preparation of fresh Chinese medicine,is officially approved by the state as a Chinese patent medicine.It consists of Xianbaihuashe(Fresh Bungarus),Xianjinshe(Fresh Agkistrodon)and Xianshougong(Fresh Gecko)and is used to treat many kinds of cancer cases.Clinical and experimental studies have shown that JLC can inhibit the growth of many malignant tumors and induce their apoptosis.Its anti-tumor effects include improving the immune system,joint intervention,radiotherapy and chemotherapy,and improving the quality of life.In this paper,the effects of JLC on proliferation and apoptosis of human gastric cancer MGC-803 and BGC-823 cells were discussed.The results of these experiments showed that JLC promoted the apoptosis of tumor cells in a certain range compared with the control group in vivo and in vitro.This study is divided into three parts.Part ? Effect of Jinlong Capsule on proliferation and apoptosis of human gastric cancer MGC-803 and BGC-823 cell LinesObjective:To study the effects of Jinlong Capsule(JLC)on proliferation and apoptosis of human gastric cancer MGC-803 and BGC-823 cells.Methods:Logarithmic human gastric cancer MGC-803 and BGC-823 cells were selected and divided into JLC concentration group(0.1,0.2,0.4,0.8 mg/ml),negative control group and positive control group(5-FU 32 ?.g/ml).MTT assay was used to detect the effects of JLC on the proliferation of human gastric cancer MGC-803 and BGC-823 cells;Electron microscopy was used to observe the changes of cell morphology and structure;Flow cytometry was used to detect the apoptotic changes of human gastric cancer MGC-803 and BGC-823 cells;Cell cycle experiments showed the changes of cell cycle after different JLC groups.Results:JLC could promote apoptosis of MGC-803 and BGC-823 cells,block cell cycle in S and G2/M phase.At the same time,JLC inhibited proliferation in a concentration-dependent manner(p<0.05 orp><0.01).Conclusion:JLC promotes apoptosis of MGC-803 and BGC-823 cells,affects cell cycle,and effectively inhibits proliferation.Part ? The molecular mechanism of Jinlong Capsule on human gastric cancer MGC-803 and BGC-823 cell linesObjective:To investigate the effects of Jinlong Capsule(JLC)on the levels of Caspase-3,Bax,Bcl-2 and Survivin in human gastric cancer MGC-803 and BGC-823 cells,and to preliminarily study the molecular mechanism of JLC inhibiting the metastasis of gastric cancer.Methods:The expression of target proteins(including Caspase-3,Bax,Bcl-2 and Survivin)in human gastric cancer MGC-803 and BGC-823 cells was detected by western blot after 24 hours of treatment with JLC(0.1,0.2,0.4,0.8 mg/ml)and 5-FU(32 ?g/ml).Results:For MGC-803 and BGC-823 cells,JLC could decrease the expression levels of Bcl-2 and Survivin protein in a concentration-dependent manner,while the expression of Bax and Caspase-3 protein increased partially with the increase of JLC concentration.Conclusion:The molecular mechanism of JLC inhibiting human gastric cancer MGC-803 and BGC-823 cells may be related to apoptosis-related proteins and Bcl-2 family.Part ? Effect of Jinlong Capsule on the growth of gastric cancer subcutaneously transplanted in nude mice and its related mechanismsObjective:To observe the effects of Jinlong Capsule(JLC)on the growth of subcutaneous transplanted gastric cancer in nude mice,and to explore whether apoptosis-related signaling pathways are involved in the inhibition of JLC on subcutaneous transplanted gastric cancer in nude mice.Methods:BGC-823 red fluorescent cells were injected subcutaneously into nude mice to establish a subcutaneous transplantation model of gastric cancer in nude mice.They were randomly divided into normal group,control group(saline solution),JLC low(390 mg/kg)?moderate(780 mg/kg),high(1560 mg/kg)dose group and 5-FU(10 mg/kg)group.All groups were given intragastric administration.The general situation of nude mice in each group was observed every day during the experiment.For the model of subcutaneous transplanted tumors,the volume of tumors was measured every two days,and the dynamic growth curve was drawn.The size of tumors was quantitatively analyzed by real-time fluorescence imaging in vivo.At the end of treatment,the nude mice in each group were executed and weighed after peeling the tumors,and the histopathological changes of tumors were observed.The expression levels of apoptotic related proteins(Bax?Bcl-2,Caspase-3 and Survivin)in tumor tissues of nude mice were detected by immunohistochemistry.Results:Subcutaneous transplantation of gastric cancer in nude mice was successfully established,and different doses of JLC was used to treat nude mice bearing gastric cancer.We found that there were no significant abnormalities in diet,behavior and mental state of nude mice in the subcutaneous transplantation model during the treatment.There was no significant difference in body weight of nude mice between the JLC groups and the control group during the whole treatment(p>0.05).The volume and weight of transplanted tumors were significantly inhibited by JLC in dose-dependent manner(p<0.01).Immunohistochemical results showed that JLC could significantly up-regulate the expression of Bax and Caspase-3 in transplanted tumors(p<0.05),and down-regulate the expression of Bcl-2 and Survivin protein(p<0.05)in a dose-dependent manner.Conclusion:JLC can inhibit the growth of nude mice bearing gastric cancer,and its mechanism may be related to the regulation of apoptotic related proteins(Bax,Bcl-2?Caspase-3 and Survivin).