A Study Of The Expression And Mechanism Of ATF2 In Esophageal Cancer

BackgroundEsophageal cancer mainly includes esophageal squamous cell carcinoma and esophageal adenocarcinoma.It is mainly caused by esophageal squamous cell carcinoma in China.China is one of the countries with high incidence of esophageal cancer in the world,and it is also one of the high mortality rates of esophageal cancer in the world.With the rapid development of medical technology in China Development,early diagnosis of esophageal cancer is greatly improved,early esophageal cancer is found in time and the prognosis of surgery is good.The 5-year postoperative survival rate can reach more than 90%.The prognosis of esophageal cancer is mostly related to the lack of early diagnosis.Most patients with esophageal cancer have symptoms.After the progression to the advanced stage,the prognosis is very poor,and the 5-year survival rate is less than 20%.In recent years,China’s research on tumor diagnosis and treatment has made great progress,but we have no effective means for the treatment of most tumors.At present,research on gene transcription regulation of tumors has made gratifying results.Activation of transcription factor-2(ATF-2)is an important transcription factor in the growth and survival of cells.Recent studies have shown that ATF2 plays an important role in the transcriptional regulation of tumor cell genes,and ATF2 is localized in cells.Different,play a role is also inconsistent,ATF2 nuclear localization and carcinogenesis,cytoplasmic localization and tumor suppression.This study investigated the role of ATF2 in esophageal cancer by detecting the expression of ATF2 mRNA and ATF2 in esophageal carcinoma and changing the expression of ATF2 on esophageal cancer cell proliferation,cell cycle and apoptosis.ObjectiveBy detecting the expression of ATF2 mRNA and ATF2 in esophageal cancer tissues,constructing a lentiviral vector overexpressing ATF2 gene,packaging lentivirus and transfecting esophageal cancer cell line to obtain a stable transgenic cell line expressing ATF2,and overexpressing ATF2 for esophageal cancer The effects of cell proliferation,cell cycle and apoptosis,explore the role of ATF2 in esophageal cancer,and provide more theoretical basis for targeted therapy of esophageal cancer.Methods1.ATF2 mRNA gene sequencing analysis was performed on esophageal cancer tissues and paracancerous tissues of 120 patients with esophageal cancer.2.Immunohistochemical staining was used to detect the expression of ATF2 protein in esophageal cancer tissues and adjacent tissues.3.Western Blot was used to detect the expression of ATF2 in 10 esophageal cancer cell lines.4.ATF2 gene fragment was obtained by PCR technique based on ATF2 gene sequence,and the lentiviral plasmid vector was packaged to obtain a lentiviral vector containing the ATF2 wild type lentiviral vector and the ATF2 Thr71 site mutation.The KYSE-140 esophageal cancer cell line was then subjected to lentiviral vector transfection experiments,and the cells were divided into 3 groups.(1)Blank control group: Transfected esophageal cancer KYSE-140 cell line with virus empty vector;(2)ATF2 wild Overexpression group: transfected esophageal cancer KYSE-140 cell line with ATF2 wild type lentiviral vector;(3)ATF2 T71 A mutant overexpression group: transfected esophageal cancer KYSE-140 cells with lentiviral vector containing ATF2 T71 A mutation A line in which the ATF2 protein expressed by the ATF2 mutant gene is a protein with impaired transcriptional activity.5.Western Blot was used to detect the expression of ATF2 protein in three cell lines after virus transfection.6.Dynamic viability survival analysis of three groups of esophageal cancer cell lines using IncuCyte Zoom,a live cell dynamic imaging and analysis system.7.Using the Annexin-FITC-PI apoptosis kit to treat three groups of esophageal cancer cells,the effect of over-expressing ATF2 cell cycle and apoptosis was detected by flow cytometry;the flow of three groups of esophageal cancer cells was treated by PI staining.The cytometer analyzed the effect of overexpression of ATF2 on the cell cycle.Results1.120 mRNA samples from patients with esophageal cancer: The expression of ATF2 mRNA in esophageal cancer tissues was higher than that in paracancerous tissues(P<0.05).2.Immunohistochemistry: The expression of ATF2 protein in esophageal cancer tissues was higher than that in paracancerous tissues(P<0.05).3.Western Blot was used to detect the expression of ATF2 in esophageal cancer cell lines: ATF2 protein was expressed in 10 esophageal cancer cell lines and was low in cell line KYSE-140.4.Western Blot was used to detect the expression of ATF2 in three groups of KYSE-140 cell lines: ATF2 expression was less in blank virus vector transfected cells,and ETF cells were transfected with ATF2 wild type lentiviral vector and ATF2 Thr71 A mutant lentiviral vector.ATF2 was overexpressed and confirmed stable transfection.5.After viral vector transfection of esophageal cancer cells,dynamic proliferation analysis of three groups of esophageal cancer cell lines: the cell proliferation level of the blank control group was higher than that of the mutant group and the wild group overexpressing ATF2(P<0.05),and the cell proliferation of the ATF2 mutant group was overexpressed.The level was higher than that of the overexpressed ATF2 wild group(P<0.05).6.Flow cytometry was used to detect the changes of G1 phase,S phase and G2 phase in the cell cycle of the three groups: the ratio of G2 and S phase in ATF2 wildtype overexpression group was higher than that in blank virus vector transfection group and ATF2 mutant overexpression group,G1 The period was lower than the blank virus vector transfection group and the ATF2 mutant overexpression group(P<0.05).The G1 phase of ATF2 mutant overexpression group was higher than that of blank virus vector transfection group and ATF2 wild type overexpression group,S phase was lower than blank virus vector transfection group and ATF2 wild type overexpression group(P<0.05),ATF2 mutant type.There was no statistically significant difference between the overexpression group and the blank virus vector transfection group(P>0.05).7.Flow cytometry was used to detect apoptosis.The apoptotic rate of ATF2 wildtype overexpressing group was significantly higher than that of blank control group and ATF2 mutant overexpression group(P<0.05).Conclusions 1.Both ATF2 mRNA and ATF2 protein were elevated in esophageal cancer tissues.2.ATF2 can promote the apoptosis of esophageal cancer cells,but also promote the cell cycle from G1 to S and G2,which is a combination of inhibition of esophageal cancer cell growth.