Research Of Celastrus Orbiculatus Extract On Inducing Mitochondrial Pathway Mediated Cell Apoptosis In Gastric Cancer Cells

Gastric cancer(GC)is one of the most deadly malignant tumors in the world,China is also a high incidence of gastric cancer.Despite significant improvements in surgical treatment and adjuvant therapy in recent decades,long-term survival remains unsatisfactory due to postoperative recurrence and metastasis.In recent 10 years,many experimental studies have proved that apoptosis is a necessary physiological process of cell death.And inducing apoptosis of cancer cells is a good strategy to treat cancer.Apoptosis is actually a kind of programmed death,mainly dependent on ATP.Apoptosis is also a self-protection mechanism of the body.The body removes unnecessary or unhealthy cells from the system in order to maintain a constant number of cells.Dysregulation of apoptosis can lead to many diseases,especially cancer.The process of cell apoptosis is multi-gene joint regulation.At present,it is believed that there are three main apoptotic signaling pathways.They are mitochondrial mediated cell apoptosis pathway,endoplasmic reticulum stress mediated cell apoptosis signaling pathway and death receptor-mediated cell apoptosis pathway.The three influence each other and constitute a regulatory network.In this paper,we mainly studied the mitochondrial mediated apoptotic signaling pathway.China is rich in traditional Chinese medicine resources,with a long history of application and clinical experience.Celastrus orbiculatus extracts(COE)is an important Chinese herbal medicine in China.Our preliminary research found that COE has significant anti-tumor activity.It can promote apoptosis and inhibit the proliferation,invasion,metastasis and angiogenesis in various malignant tumor cells.The main purpose of this study is to investigate the effect of COE on the proliferation and apoptosis of SGC-7901 and MKN45 cells and to explore the molecular mechanism induced by COE.The results showed that COE could promote the apoptosis of SGC-7901 and MKN45 cells,and the molecular mechanism of promoting the apoptosis of gastric cancer cells was related to mitochondrial metabolismThis study is divided into two partsPart ?Effect of Celastrus Orbiculatus Extract on proliferation and apoptosis of human gastric cancer MKN45 and SGC-7901 cellsObjective:To study the effects of Celastrus orbiculatus extracts(COE)on proliferation and apoptosis of human gastric cancer MKN45 and SGC-7901 cells.Methods:Logarithmic human gastric cancer MKN45 and SGC-7901 cells were selected and divided into COE concentration group(10,20,40,80,160,320mg/L),negative control group and positive control group(5-Fu 32mg/L).MTT assay was used to detect the effects of COE on the proliferation of human gastric cancer MKN45 and SGC-7901 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 MKN45 and SGC-7901 cells.Results:COE could promote the apoptosis of MKN45 and SGC-7901 cells and inhibit cell proliferation in a concentration-dependent manner(*p<0.05,**p<0.01),COE could destroy the ultrastructure of human gastric cancer cellsConclusion:COE promotes apoptosis of MKN45 and SGC-7901 cells and inhibits proliferation effectivelyPart ?The molecular mechanism of apoptosis induced by COE in MKN45 and SGC-7901 cellsObjective:To investigate the effect of Celastrus orbiculatus extracts(COE)on the function of mitochondria and the expression of related proteins in MKN45 and SGC-7901 cells,and to explore the possible molecular mechanism of inhibiting the occurrence and development of gastric cancer.Methods:Different concentrations of COE(20,40,80 mg/L)and 5-Fu(32 mg/L)were added to logarithmic human gastric cancer MKN45 and SGC-7901 cells.After 24 hours of treatment,the changes of mitochondrial membrane potential were observed by fluorescence microscope.Electron microscopy was used to observe the changes of mitochondrial structure in gastric cancer cells MKN45 and SGC-7901 treated with COE(80mg/L).With Mito-tracker Red staining was used to observe changes in mitochondrial mass with confocal microscope,flow cytometry was used to detect the content of total ROS in cells,and Western blot was used to detect the expression levels of related proteins(Bax,Bcl-2,Cyt-C,Caspase-3 and Caspase-9)Results:For MKN45 and MKN45 cells,COE could decrease the mitochondrial membrane and mitochondrial mass,and increase the level of ROS.Compared with the wild group,the disorder of ridge arrangement was not obvious in the COE group.COE could decrease the protein expression level of Bcl-2 in gastric cancer cells,while the protein expression of Bax,Cyt-C,Caspase-3,Caspase-9 increased with the increase of COE concentrationConclusion:The molecular mechanism of COE inducing apoptosis of MKN45 and SGC-7901 cells may be related to mitochondrial pathway.