Objective The aim of the study was to investigate the effects of Chinese herbal formula Weichang’an(WCA)on the proliferation and mitochondria-mediated apoptosis of human gastric cancer cells.Methods Cell Counting Kit...Objective The aim of the study was to investigate the effects of Chinese herbal formula Weichang’an(WCA)on the proliferation and mitochondria-mediated apoptosis of human gastric cancer cells.Methods Cell Counting Kit-8(CCK8)was used to evaluate the antiproliferative activity ofWCA on MKN45 cells;Giemsa staining was used to investigate cell colony formation;flow cytometry was used to analyze cell cycle,apoptosis rate,and caspases activation;and Hoechst staining was used to analyze the morphology of cell nuclei.The mitochondrial membrane potential was analyzed by both flow cytometric measurements and fluorescence microscopy.Western blot was used to analyze the protein levels of pro-caspase-3,Bcl-2,Bax,and Bcl-X.MKN45 cells were subcutaneously injected into the right forelimb of 16 nude mice to establish the tumor xenograft model,and a total of 12 nude mouse tumor xenograft models were successfully created.The nude mice were divided into the control group and the WCA-treated group.The two groups received normal saline and WCA treatment at 35.49 g/kg by a single daily oral gavage for 21 days.The body weight and tumor size of the nude mice were measured twice a week.The ultrastructural changes of subcutaneous tumors were observed by a transmission electron microscope.Results WCA can suppress cell proliferation and colony formation.It can also induce changes in the mitochondrial membrane potential and increase the activities of caspases-3,caspases-8,and caspases-9 in MKN45 cells.WCA induced S-phase arrest and apoptosis in MKN45 cells,and decreased the expression levels of antiapoptotic proteins Bcl-2 and Bcl-X in MKN45 cells,while increasing the expression levels of the proapoptotic proteins Bax and pro-caspase-3 compared with the control group.WCA inhibited the growth of a xenografted MKN45 tumor in nude mice,and the protein levels of Bax and pro-caspase-3 were significantly increased,while Bcl-X and Bcl-2 were reduced compared with the control group.The differences are statistically significant(p<0.05).Conclusion WCA could suppress the proliferation of gastric cancer cells via mitochondrial apoptosis.展开更多
Objective The aim of the study was to screen key proteins involved in the treatment of human gastric cancer subcutaneous xenografts in nude mice by the traditional Chinese medicine formula Weichang’an and to explore ...Objective The aim of the study was to screen key proteins involved in the treatment of human gastric cancer subcutaneous xenografts in nude mice by the traditional Chinese medicine formula Weichang’an and to explore its mechanism of action in treating gastric cancer.Methods Sixteen 7-to 8-week-old female BALB/C nude mice were used to establish a human gastric cancer subcutaneous xenograft model by bilateral axillary injection of MKN45 cells.Mice with successfully established tumors were randomly divided into the Weichang’an group and the model group,with eight mice in each group.Mice in the Weichang’an group were orally administered 0.5mL of Weichang’an decoction,while mice in the model group were given 0.5 mL of normal saline by gavage once a day for 21 consecutive days.On day 28,the animals were sacrificed by cervical dislocation,and tumors were excised to measure tumor weight and assess the tumor suppression rate.Tandem mass tags(TMT)quantitative proteomics was used to analyze the tumor samples,identify differentially expressed proteins,and perform Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Results Compared with the model group,the tumor weight in the Weichang’an group was significantly reduced(p<0.001),and the tumor suppression rate was 41.40%,indicating that Weichang’an can inhibit the growth of human gastric cancer subcutaneous xenografts in nude mice.TMT quantitative proteomics identified a total of 2,856 proteins,with 13 proteins showing downregulated expression and 25 proteins showing upregulated expression in the Weichang’an group.GO enrichment analysis revealed that differentially expressed proteins were mainly enriched in cellular components such as the cell membrane,extracellular matrix,and envelope,and participated in biological processes like negative regulation of cell adhesion hematopoiesis,and skeletal system development,with functions in cell adhesion molecule binding and molecular sensor activity.The KEGG pathway enrichment analysis identified the autophagy-lysosome pathway.Conclusion Weichang’an may exert its therapeutic effect on gastric cancer by regulating the expression of various proteins and modulating the autophagy-lysosome pathway.展开更多
基金funded by the Three-Year Plan of Action for the Development of Traditional Chinese Medicine in Shanghai(ZY3-CCCX-3-2003)State Administration of Traditional Chinese Medicine(TCM)Clinical Research Base of China(JDZX2015068)+1 种基金the Key and Promotion Special Topics of Henan(202102310164)Henan Province Traditional Chinese Medicine Scientific Research Special Project(2022ZY2031).
文摘Objective The aim of the study was to investigate the effects of Chinese herbal formula Weichang’an(WCA)on the proliferation and mitochondria-mediated apoptosis of human gastric cancer cells.Methods Cell Counting Kit-8(CCK8)was used to evaluate the antiproliferative activity ofWCA on MKN45 cells;Giemsa staining was used to investigate cell colony formation;flow cytometry was used to analyze cell cycle,apoptosis rate,and caspases activation;and Hoechst staining was used to analyze the morphology of cell nuclei.The mitochondrial membrane potential was analyzed by both flow cytometric measurements and fluorescence microscopy.Western blot was used to analyze the protein levels of pro-caspase-3,Bcl-2,Bax,and Bcl-X.MKN45 cells were subcutaneously injected into the right forelimb of 16 nude mice to establish the tumor xenograft model,and a total of 12 nude mouse tumor xenograft models were successfully created.The nude mice were divided into the control group and the WCA-treated group.The two groups received normal saline and WCA treatment at 35.49 g/kg by a single daily oral gavage for 21 days.The body weight and tumor size of the nude mice were measured twice a week.The ultrastructural changes of subcutaneous tumors were observed by a transmission electron microscope.Results WCA can suppress cell proliferation and colony formation.It can also induce changes in the mitochondrial membrane potential and increase the activities of caspases-3,caspases-8,and caspases-9 in MKN45 cells.WCA induced S-phase arrest and apoptosis in MKN45 cells,and decreased the expression levels of antiapoptotic proteins Bcl-2 and Bcl-X in MKN45 cells,while increasing the expression levels of the proapoptotic proteins Bax and pro-caspase-3 compared with the control group.WCA inhibited the growth of a xenografted MKN45 tumor in nude mice,and the protein levels of Bax and pro-caspase-3 were significantly increased,while Bcl-X and Bcl-2 were reduced compared with the control group.The differences are statistically significant(p<0.05).Conclusion WCA could suppress the proliferation of gastric cancer cells via mitochondrial apoptosis.
基金funded by the Special Research Project of Traditional Chinese Medicine in Henan Province(2022ZY2031).
文摘Objective The aim of the study was to screen key proteins involved in the treatment of human gastric cancer subcutaneous xenografts in nude mice by the traditional Chinese medicine formula Weichang’an and to explore its mechanism of action in treating gastric cancer.Methods Sixteen 7-to 8-week-old female BALB/C nude mice were used to establish a human gastric cancer subcutaneous xenograft model by bilateral axillary injection of MKN45 cells.Mice with successfully established tumors were randomly divided into the Weichang’an group and the model group,with eight mice in each group.Mice in the Weichang’an group were orally administered 0.5mL of Weichang’an decoction,while mice in the model group were given 0.5 mL of normal saline by gavage once a day for 21 consecutive days.On day 28,the animals were sacrificed by cervical dislocation,and tumors were excised to measure tumor weight and assess the tumor suppression rate.Tandem mass tags(TMT)quantitative proteomics was used to analyze the tumor samples,identify differentially expressed proteins,and perform Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Results Compared with the model group,the tumor weight in the Weichang’an group was significantly reduced(p<0.001),and the tumor suppression rate was 41.40%,indicating that Weichang’an can inhibit the growth of human gastric cancer subcutaneous xenografts in nude mice.TMT quantitative proteomics identified a total of 2,856 proteins,with 13 proteins showing downregulated expression and 25 proteins showing upregulated expression in the Weichang’an group.GO enrichment analysis revealed that differentially expressed proteins were mainly enriched in cellular components such as the cell membrane,extracellular matrix,and envelope,and participated in biological processes like negative regulation of cell adhesion hematopoiesis,and skeletal system development,with functions in cell adhesion molecule binding and molecular sensor activity.The KEGG pathway enrichment analysis identified the autophagy-lysosome pathway.Conclusion Weichang’an may exert its therapeutic effect on gastric cancer by regulating the expression of various proteins and modulating the autophagy-lysosome pathway.
