Experimental Study On The Effects Of Compound Lizard Powder Different Microparticle Combination On Proliferation,apoptosis And SIRT1,P53 Of Human Gastric Cancer Cell SGC-7901

ObjectiveTo observe the effects of compound Lizard Powder on the proliferation and apoptosis of human SGC-7901 gastric cancer cells and the expression of SIRT1 and P53protein,and to elucidate the mechanism of the intervention of gastric cancer cells.Methods160 SPF male Sprague-Dawley rats were randomly divided into 8 groups:normal saline group,compound lizard powder 80 mesh group,100 mesh group,80 mesh+100mesh mixed high dose group,middle dose group,low dose group,Huachansu tablets group,5-Fu group,20 in each group.Each group was given intragastric or tail vein injection for 1 week,and the drug-containing serum was prepared by centrifugation of abdominal aorta in a sterile environment,and then added to human SGC-7901 gastric cancer cells for a predetermined time.The proliferation activity of SGC-7901 cells cultured in vitro by compound rat lizard powder was determined by MTT assay.Annexin V-FITC/PI double staining and flow cytometry were used to detect the effect of compound lizard powder on apoptosis and cycle of SGC-7901 cells.The effects of compound lizard powder on the expression of SIRT1 and P53 protein in SGC-7901 cells were detected by immunocytochemistry and Western-Blot assay.Statistical analysis was performed using SPSS20.0 For Windows software.Results1.Cell proliferation was detected by MTT:Compared with the saline group,the cell proliferation of each drug serum group was significantly inhibited（P<0.05）.Compared with the 5-Fu group and the Huachansu tablets group,the OD mean of each Chinese medicine group decreased（P<0.05）.The OD value of 80 mesh+100 mesh mixed group was less than 80mesh group and 100 mesh group（P<0.05）.The mixed groups were compared with each other,and the high dose group with 80 mesh+100 mesh was the best（P<0.05）.2.The effect of Annexin V-FITC/PI double staining on apoptosis:Compared with the saline group,the total apoptosis rate of the cells in the serum intervention group was significantly increased（P<0.05）.The total apoptotic rate of the cells in each Chinese medicine group was greater than that of 5-Fu group and Huachansu tablets group（P<0.05）.The total apoptotic rate of the 80 mesh+100 mesh mixed group was greater than 80 mesh group and100 mesh group（P<0.05）.The mixed groups were compared with each other,and the total apoptotic rate was the highest in the high-dose group with 80 mesh+100 mesh（P<0.05）.3.The cell cycle was detected by flow cytometry:Compared with the saline group,the proportion of cells in the G₁ phase of each drug serum intervention group was significantly increased（P<0.05）.The proportion of cells in G₁ phase of each Chinese medicine group was higher than that of Huachansu tablets group and 5-Fu group（P<0.05）.The proportion of G₁phase cells in the 80 mesh+100 mesh mixed group was greater than that in the 80 mesh group and the 100 mesh group（P<0.05）.The ratio of G₁ phase cells in the high dose group mixed with 80 mesh+100 mesh was the largest among the mixed groups（P<0.05）.4.The expression of SIRT1 and P53 in human SGC-7901 gastric cancer cells was detected by immunocytochemistry:Compared with the saline group,the positive expression of SIRT1 and P53 protein in each drug-containing serum group decreased to different degrees（P<0.05）.The positive expressions of SIRT1 and P53 protein in the Chinese medicine group were lower than those in the Huachansu tablets group and 5-Fu group（P<0.05）.The positive expression of SIRT1 and P53 protein in the 80 mesh+100 mesh mixed group was lower than that in the 80 mesh group and the 100 mesh group（P<0.05）.Comparison of each mixed group,the positive expression of SIRT1 and P53 protein was the lowest in the 80+100mesh high dose group（P<0.05）.5.The expression levels of SIRT1 and P53 in human SGC-7901 gastric cancer cells were detected by Western-Blot:Compared with the saline group,the gray values of SIRT1 and P53in each drug-containing serum group were decreased to different degrees（P<0.05）.The gray values of SIRT1 and P53 in each Chinese medicine group were lower than those in Huachansu tablets group and 5-Fu group（P<0.05）.The gray values of SIRT1 and P53 in the 80mesh+100 mesh mixed group were lower than those in the 80 mesh group and the 100 mesh group（P<0.05）.For each mixed group,the gray values of SIRT1 and P53 were the lowest in the80 mesh+100 mesh high dose group（P<0.05）.Conclusion1.Compound Lizard Powder Different Particle Combination Agent can inhibit the proliferation of human SGC-7901 gastric cancer cells and induce apoptosis.2.Compound Lizard Powder Different Particle Combination Agent can promote cell cycle arrest in human SGC-7901 gastric cancer and arrest cells in G₁ phase.3.Compound Lizard Powder Different Particle Combinations can reduce the expression of SIRT1 and P53 proteins,inhibit the canceration by inhibiting SIRT1-mediated P53signaling pathway,and play a role in inhibiting tumor growth.