Study Of Naa10P On The Cycle Of Esophageal Squamous Cell Carcinoma Cells And Its Sensitivity To Cisplatin

Objective: Esophageal Squamous-Cell Carcinoma(ESCC),the expression level of N-α acetyltransferase(Naa10P)in cancer tissues is significantly higher than that in adjacent tissues.Esophageal cancer is a serious threat to human health.Squamous cell carcinoma is more common in the pathological classification of malignant tumors.The prognosis of patients is extremely poor,their 5-year survival rate is low,and patients often have chemotherapy resistance.At present,there is no effective targeted drug to inhibit the growth of esophageal cancer cells,and the specific mechanism involved in regulating the proliferation cycle of esophageal cancer cells is still unclear.Due to the high expression of Naa10 P in esophageal cancer tissues,this study used the in vitro culture of the esophageal squamous cell carcinoma ECA109 cell line to explore the effect of Naa10 P expression on the cell cycle and drug sensitivity of esophageal squamous cell carcinoma,and explained that Naa10 P passes through β-Catenin signaling pathway regulates its cell cycle and determines the influence of Naa10 P expression on the sensitivity of esophageal squamous cell carcinoma cells to the chemotherapy drug cisplatin(DDP),Provide new ideas in the treatment of esophageal squamous cell carcinoma and increase the sensitivity of chemotherapy drugs.Method:1.Use small interfering RNA(siRNA)to construct a human esophageal squamous cell carcinoma ECA109 cell line with low Naa10 p expression.The experiment is divided into ECA109-siRNA-Naa10 group,ECA109-siRNA-NC group,and ECA109-blank group.2.Identify the interference efficiency of siRNA on cells by fluorescence microscopy,qrt-PCR and Western blotting.3.Flow cytometry to detect cell cycle,CCK-8 method to draw cell growth curve,Western blot method to identify β-Catenin,acetylated β-Catenin(Acetyl-β-Catenin),Cyclin D1,C-The protein expression level of Myc.4.Use the CCK-8 method to detect the sensitivity of each group of cells to DDP after treatment with different DDP concentrations.Results:1.siRNA transfection and transfection efficiencySet different Lip2000 and siRNA concentration ratios,observe with fluorescence microscope,and give the best transfection ratio when the concentration ratio of each well(six-well plate)is 10 u L:10u L.The successful transfection was identified by qrt-PCR and Western blotting,and the difference was statistically significant(P <0.01).2.Western blotting to identify the expression levels of key proteins in the β-Catenin signaling pathwayThe expression levels of β-Catenin in ECA109-siRNA-Naa10 group,ECA109-siRNA-NC group,and ECA109-blank group were not different(P>0.05).The acetylated β-Catenin,Cyclin D1,C-Myc of ECA109-siRNA-Naa10 group The expression level was significantly lower than the ECA109-siRNA-NC group and ECA109-blank group,the difference was statistically significant(P <0.05).3.Flow cytometry to detect cell cycleThe G0/G1 phase of the cell cycle detected by flow cytometry showed: ECA109-siRNA-Naa10 experimental group(64.833±5.685),ECA109-siRNA-NC control group(56.833±1.793),ECA109-blank group(54.700±4.762).The proportion of cells in the G0/G1 phase of the ECA109-siRNA-Naa10 experimental group was significantly increased compared with the other two groups,and the difference was statistically significant(P<0.05).4.CCK-8 method to draw cell growth curveThe cell cycle growth curve of ECA109-siRNA-Naa10 group was slower than that of ECA109-siRNA-NC group and ECA109-blank group(P<0.01),that is,cell proliferation decreased.5.Use the CCK-8 method to detect the sensitivity of each group of cells to DDP after treatment with different DDP concentrations.The IC50 of ECA109-siRNA-Naa10 group,ECA109-siRNA-NC group and ECA109-blank group on DDP were(12.776±0.739)μg/m L,(19.223±0.276)μg/m L,(18.526)±0.568)μg/m L,the difference was statistically significant(P <0.01).Conclusion:1.By reducing the expression of Naa10 p,the expression standard of acetylated β-catenin,target genes Cyclin D1 and C-Myc in the β-catenin signaling pathway are obviously reduced,the esophageal squamous cell carcinoma cell cycle is blocked in G0/G1 section,and cell proliferation is significantly reduced slow.2.By reducing the expression of Naa10 p,the IC50 value of esophageal squamous cell carcinoma cell lines is significantly reduced,which can increase the sensitivity of ECA109 cells to DDP.