| Compound Cantharide Injection（CCI）is a combination of the traditional Chinese medicine cantharidin for promoting blood circulation and removing blood stasis and Fuzhengliqi traditional Chinese medicine ginseng,astragalus and acanthopanax.It has beneficial effects of nourishing qi and nourishing yin,clearing heat and detoxifying,eliminating blood stasis and dissolving stagnation,and has a good therapeutic effect on human liver cancer,lung cancer,rectal cancer,etc.It is widely used in clinical treatment of tumors,but the compound research of compound canthari soup on gastric cancer And the corresponding mechanism of action is less studied,and the related mechanism is not clear.In the early stage of this research group,the network pharmacology and the method of big data were used to analyze and predict the mechanism of action of Fufang Bandao Decoction.Based on this,the anti-gastric cancer mechanism of Fufang Bandao Decoction predicted by network pharmacology is experimentally verified.In this thesis,human gastric cancer SGC-7901 cells are used as the research object.First,the pharmacodynamics of compound canthari decoction to inhibit the proliferation of human gastric cancer SGC-7901 cells were investigated,and then the mechanism of human gastric cancer SGC-7901cells induced cycle arrest was investigated.Research was conducted,and finally the related protein pathways that inhibit the proliferation of gastric cancer cells were verified.This article mainly carried out five aspects of research,namely:（1）the effect of compound canthari soup on human gastric cancer SGC-7901 cell cytotoxicity;（2）the effect of compound canthari soup on human gastric cancer SGC-7901 cell cycle block;（3）the effect of compound canthari soup on gastric cancer SGC-7901 cells TNF-αpathway;（4）the effect of compound canthari soup on gastric cancer SGC-7901 cells AR pathway;（5）the compound canthari soup on gastric cancer SGC-7901 cells TGF-βpathway effects.In the study of the cytotoxic effect of compound canthari soup on human gastric cancer SGC-7901 cells,the MTT method and the method of drawing cell growth curves have been used to observe the inhibitory effect of compound canthari soup on human gastric cancer SGC-7901 cells.SGC-7901 cells have a good proliferation inhibition effect.Inverted microscope and fluorescence microscope were used to observe the effect of Fufang Bandao Decoction on the morphology of human gastric cancer SGC-7901 cells.It was found that Fufang Bandao Decoction can significantly change the morphology of human gastric cancer SGC-7901 cells,destroy the cell membrane and nuclear structure and promote the death of human gastric cancer SGC-7901 cells..Through this part of research,it is shown that the compound canthari soup has a significant inhibitory effect on the proliferation of human gastric cancer SGC-7901 cells.The results of network pharmacology research show that the inhibition of compound canthari decoction on human gastric cancer can cause human gastric cancer cell cycle arrest,regulate human gastric cancer cell TNF-αsignaling pathway,AR signaling pathway and TGF-βsignaling pathway,so we next The effects of Fufang Banzhi Decoction on human gastric cancer SGC-7901 cell cycle,TNF-αsignal pathway,AR signal pathway and TGF-βsignal pathway were observed.In the study of compound canthari decoction on cell cycle arrest of human gastric cancer SGC-7901,the effects of compound canthari decoction on the cell cycle of human gastric cancer SGC-7901 were determined by flow cytometry and PI single staining.SGC-7901 cells were arrested in the S phase;the effect of Fufang Banzhi Decoction on the expression of cell cycle regulation-related proteins in human gastric cancer SGC-7901 was detected,and it was found that the expression of CDK4 and cyclinD1 showed an increasing trend with the increase of drug concentration,prompting the presence of G0/G1cells gradually changed to S-phase cells,and the expression of CDK2 and cyclin A showed a downward trend,which prevented S-phase cells from entering G2phase.Among the effects of compound canthari decoction on human gastric cancer SGC-7901 cells,the effect of compound canthari decoction on TNF-α,IKB-α,NF-κB,MEKK1,MKK7,JNK,caspase 8,and The effects of caspase 3 and caspase 7 protein expressions were found to increase TNF-α,P-IKBα,MEKK1,P-MKK7,P-JNK,cleaved caspase 8,cleaved caspase 3,and cleaved caspase as the concentration of compound canthari soup increased 7 down-regulates the expression of IKB-α,NF-κB,JNK and caspase 3,but has no significant effect on the expression of three proteins,MKK7,caspase 8 and caspase 7,indicating that compound canthari decoction can regulate human gastric cancer SGC-7901 cells The caspase family-mediated apoptosis pathway,the NF-κB signaling pathway involved in transcriptional regulation,and the JNK signaling pathway are three classic TNF-αpathways that inhibit the proliferation of human gastric cancer SGC-7901 cells.In the process of the effect of compound canthari decoction on the AR pathway of human gastric cancer SGC-7901 cells,the effect of compound canthari decoction on the expression of AR and STEAP1 in human gastric cancer SGC-7901 cells was detected.The expressions of AR（nucleus）and STEAP1 proteins indicate that the compound canthari soup promotes the distribution of AR protein in cytoplasm and nucleus of human gastric cancer SGC-7901 cells and regulates the AR protein pathway to inhibit the proliferation of human gastric cancer SGC-7901 cells.Finally,the effect of compound canthari decoction on human gastric cancer SGC-7901 cells TGF-βprotein pathway was studied.The effect of compound canthari decoction on the expression levels of TGF-β,smad2/3 and smad4 of gastric cancer SGC-7901 cells was detected.Concentration could up-regulate the expression of smad4 protein,down-regulate the expression of TGF-βand p-smad2/3 protein,but had no significant effect on smad2/3 protein expression.It is shown that the compound cantharius decoction can inhibit the proliferation of human gastric cancer SGC-7901 cells by regulating the TGF-βprotein.In this study,we found that compound canthari decoction can inhibit the proliferation of human gastric cancer SGC-7901 cells.This effect is closely related to blocking human gastric cancer SGC-7901 cells in the S phase and causing cell death.Further research has found that proliferation inhibition can regulate cell cycle related proteins,TNF-αpathway,AR pathway and TGF-βsignal pathway are related,which validates the mechanism of Fufang Bandao Decoction predicted by network pharmacology in inhibiting the proliferation of human gastric cancer. |
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