Expression And Function Of Activator Of Basal Transcription 1 In Gastric Carcinoma

ObjectiveTo investigate the expression and clinical significance of activator of basal transcription 1(ABT1)in gastric cancer and its relationship with the development of gastric cancer;To investigate the effect of ABT1 overexpression on the growth and migration of gastric cancer cells and to provide experimental evidence for early clinical diagnosis of gastric cancer.Methods1.Expression and analysis of ABT1 in gastric cancer tissues: The expression of ABT1 in 100 cases of human gastric adenocarcinoma tissue chip sample TMA and the expression of 80 cases of adjacent gastric tissues paired with gastric cancer tissue were detected by immunohistochemical SP method.The relationship between the expression level of ABT1 in gastric cancer tissue samples and the clinicopathological features and prognosis of gastric cancer was analyzed.2.Cell strain screening experiments: Human gastric epithelial cell line GES-1,moderately differentiated gastric adenocarcinoma cell line SGC-7901 and poorly differentiated gastric cancer cell line BGC-823 cells were cultured in vitro.The basic expression of ABT1 in 3 kinds of cells was detected by Western Blot.3.An overexpression plasmid pc DNA3.1-ABT1-myc was constructed.The full-length sequence of mouse ABT1 was amplified,and the construction and identification of pc DNA3.1/myc-ABT1 eukaryotic expression vector.4.Gene transfection: BGC-823 gastric cancer cells were cultured,and the effect of ABT1 gene upregulation on proliferation and migration of gastric cancer cell lines was observed.Transfection experiments were divided into untreated group,empty vector transfection group and ABT1 gene transfection group.Western blot was used to detect the expression of ABT1 protein in each group of cells.5.The proliferation of cells in each group was detected by MTT assay.6.Scratch test to detect cell migration ability in each group.Results1.Expression and significance of ABT1 in gastric cancer tissues: ABT1 is mainly distributed in the nucleus and cytoplasm of gastric cancer and adjacent tissues,and the expression of ABT1 in gastric cancer tissues is lower than that in adjacent tissues(p=0.021).There was a significant negative correlation between ABT1 protein expression and pathological grade in gastric cancer(p = 0.026).Kaplan-Meier survival curve analysis showed that patients with high expression of ABT1 were positively correlated with good prognosis(p<0.0001).In addition,the survival rate of high expression of ABT1 in gastric cancer patients was significantly higher than that of patients with low expression of ABT1(p=0.0272).2.Cell line screening: Western Blot results showed that the protein expression of ABT1 in BGC-823 cells was significantly lower than that of GES-1 and SGC-7901 cells(p<0.01),so BGC-823 cells were selected for subsequent experiments.3.ABT1 gene transfection: PCR identification and sequencing analysis showed that the recombinant plasmid pc DNA3.1-ABT1-myc was successfully constructed.After transfection of the plasmid into BGC-823 cells,the expression of ABT1 protein in the empty vector transfection group was not significantly different(p>0.05),and the ABT1 protein expression was significantly increased in the ABT1 transfection group(p < 0).01)shows successful transfection of the ABT1 gene.4.ABT1 gene up-regulates the proliferation and migration of gastric cancer cell lines: MTT results showed that the cell proliferation rate of ABT1 transfection group was significantly lower than that of empty vector control group at 24 h,48 h,and 72 h(p < 0.01).The scratch test results showed that compared with the control group,the ABT1 transfection group significantly inhibited the migration of BGC-823 cells(p < 0.01).Conclusion1.Ectopic expression of ABT1 plays an important role in the development of gastric cancer.Low expression of ABT1 was positively correlated with poor prognosis in patients with gastric cancer,while patients with high ABT1 protein expression had longer overall survival.2.Overexpression of ABT1 inhibits the proliferation and migration of human poorly differentiated BGC-823 gastric cancer cell lines,and its mechanism needs further exploration.