| Objective1.By studying the specific biological effects of Yin-nourishing,Spleen-tonic and Detoxification Prescription water extract on gastric cancer SGC-7901 cells and MGC-803 cells,it is to explore the anti-tumor mechanism of Yin-nourishing,Spleen-tonic and Detoxification Prescription from the aspects of regulating tumor cell proliferation,migration,apoptosis,cycle,angiogenesis and related gene mRNA and protein expression levels.2.To provide an objective basis for the clinical use of gastric cancer treatment,to provide new ideas for studying the mechanism of the effective components of anti-tumor traditional Chinese medicine,and to explore from point to point.MethodsYin-nourishing,Spleen-tonic and Detoxification Prescription water extract was carried out.Gastric cancer SGC-7901 cells and MGC-803 cells were cultured in vitro.The effect of Yin-nourishing,Spleen-tonic and Detoxification Prescription water extract on the proliferation of gastric cancer SGC-7901 cells and MGC-803 cells was detected by CCK8 assay.The migration ability of cells was detected by scratch method and the apoptosis was detected by flow cytometry(Annexin V-FITC and PI staining and flow cytometry analysis).RT-qPCR and Western Blot assays were used to detect the expression levels of Bax,Bcl-2,P53,Caspase3,CDK4 and Vegf genes and proteins in the two kinds of cells after the intervention of Yin-nourishing,Spleen-tonic and Detoxification Prescription water extract.Results1.Yin-nourishing,Spleen-tonic and Detoxification Prescription water extract could inhibit the proliferation of gastric cancer SGC-7901 cells and MGC-803 cells in a dose-and-time-dependent manner(P<0.01).The half-inhibition rate(IC50)of the two kinds of cells after 48 hours of taking Yin-nourishing,Spleen-tonic and Detoxification Prescription water extract was carried out for subsequent experiments.The IC50 of SGC-7901 cells was 0.55 1%,and the IC50 of MGC-803 cells was 0.506%;2.Scratch method results showed:Yin-nourishing,Spleen-tonic and Detoxification Prescription water extract could inhibit the migration of SGC-7901 cells and MGC-803 cells in a time-dependent manner(P<0.05);3.FIow cytometry(Annexin V-FITC and PI staining and flow cytometry analysis)showed:the apoptosis rate of SGC-7901 cells and MGC-803 cells in the experimental group was significantly higher than that in the control group(P<0.05);4.RT-qPCR results showed:for the experimental group SGC-7901 cells,the mRNA expression levels of Bax,Caspase3 and Vegf genes were up-regulated,and the mRNA expression levels of Bcl-2,P53 and CDK4 genes were down-regulated(P<0.05);for the experimental group MGC-803 cells,the mRNA expression levels of Bax,Caspase3,P53 and Vegf genes were up-regulated,and the mRNA expression levels of Bcl-2 and CDK4 genes were down-regulated(P<0.05).5.Western Blot results showed:for the experimental group SGC-7901 cells and MGC-803 cells,the expression levels of Bax and P53 proteins were significantly up-regulated,the expression of Pro-Caspase3 protein was slightly down-regulated,the level of Cleaved-Caspase3 protein was significantly up-regulated,and the expression levels of Bcl-2,CDK4 and Vegf proteins were significantly down-regulated(P<0.05).ConclusionYin-nourishing,Spleen-tonic and Detoxification Prescription could inhibit the proliferation of gastric cancer SGC-7901 cells and MGC-803 cells,weaken their migration ability,and promote cell apoptosis.Its anti-tumor mechanism may be related to its intervention on the expression of Bax,Bcl-2,P53,Caspase3,CDK4,Vegf genes and protein levels. |
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