Mechanism Of LncRNA GAS5/miR-21/SOX6 Signaling Axis Regulating The Progress Of Kazakh Esophageal Carcinoma In Xinjiang

Objective: By detecting the expression of lncRNA GAS5 in Xinjiang Kazakh esophageal cancer patients and analyzing its correlation with clinicopathological features,the related role of GAS5 in esophageal cancer was explored.The proliferation and apoptosis of esophageal cancer cells were detected by overexpressing GAS5.The role of invasion,migration,and further bioinformatics predictions suggest that miR-21 may be a downstream target gene of GAS5,and SOX6 may be a downstream target gene of miR-21,suggesting that GAS5 may regulate SOX6 through competitive binding to miR-21.The expression level,thus participating in the occurrence and development of esophageal cancer,provides a potential target for the diagnosis and treatment of esophageal cancer.Methods: The expression levels of GAS5 in esophageal cancer tissues and cell lines(EC9706,EC109,KYSE150 and KYSE45)of Xinjiang Kazakh nationality were detected by q PCR,and the correlation between the expression levels and clinicopathological characteristics was analyzed.MTT assay was used to detect the effect of overexpression of GAS5 on the proliferation of esophageal cancer cells.The migration and invasion of esophageal cancer cells were detected by Transwell assay.Flow cytometry was used to detect the effect of overexpression of GAS5 on apoptosis of esophageal cancer cells.he target miRNA of GAS5 were predicted by biological information,and the targeting relationship between GAS5 and miR-21 was verified by double luciferase,RIP and RNA Pull-down experiments.The expression levels of miR-21 in esophageal cancer tissues and cell lines(EC9706,EC109,KYSE150,KYSE45)of Xinjiang Kazakh nationality were detected by q PCR,and the correlation between miR-21 expression and GAS5 was analyzed.GAS5 was up-regulated or down-regulated to observe the expression of miR-21.Response experiments were conducted to investigate whether GAS5 regulates the occurrence and development of esophageal cancer through miR-21.The target m RNA of miR-21 were predicted by biological information,and the targeting relationship between miR-21 and SOX6 was verified by double luciferase and RIP experiments.The expression levels of SOX6 in esophageal cancer tissues of Xinjiang Kazakh nationality were detected by q PCR and immunohistochemistry,and the correlation between SOX6 expression and miR-21 was analyzed.The SOX6 expression in cell lines were detected by q PCR and Western blot;miR-21 was up-regulated or down-regulated to observe the expression of SOX6.Response experiments were conducted to investigate whether miR-21 regulates the occurrence and development of esophageal cancer through SOX6.Results: 1)The expression of GAS5 in esophageal cancer tissues and cell lines were significantly down-regulated(P<0.05),and correlated with TNM stage.Compared with the control group,transfected pc DNA-GAS5 significantly increased the expression of GAS5 in esophageal cancer cell lines(EC9706,EC109)(P<0.05).Overexpression of GAS5 significantly inhibited cell proliferation,invasion and migration,and induced cell apoptosis(P<0.05).2)Bioinformatics predictions show that miR-21 is a potential target gene for GAS5.Double luciferase,RIP and RNA Pull-down experiments confirmed that miR-21 was the target gene of GAS5.The expression of miR-21 in esophageal cancer tissues and cell lines was significantly up-regulated(P<0.05),and its expression was negatively correlated with GAS5.Up-regulation of GAS5 in esophageal cancer cell lines(EC9706,EC109)significantly inhibited the expression of miR-21(P<0.05),while down-regulation of GAS5 significantly promoted the expression of miR-21(P<0.05).Co-transfection of GAS5 and miR-21 vectors significantly restored the inhibition of GAS5 on the expression of miR-21,reversed the inhibition of GAS5 on the proliferation,migration and invasion of esophageal cancer cells,and saved the induction of apoptosis of GAS5 on esophageal cancer cells.3)Bioinformatics predictions show that SOX6 is a potential target gene for miR-21.Double luciferase,and RIP experiments confirmed that SOX6 was the target gene of miR-21.The expression of SOX6 in esophageal cancer tissues and cell lines was significantly down-regulated(P<0.05),and its expression was negatively correlated with miR-21.Up-regulation of miR-21 significantly inhibited the protein expression of SOX6(P<0.05),while down-regulation of miR-21 significantly promoted the protein expression of SOX6(P<0.05).Co-transfection of anti-miR-21 and si RNA-SOX6 significantly reversed the inhibition of anti-miR-21 on the proliferation,migration and invasion of esophageal cancer cells,and saved the induction of apoptosis of anti-miR-21 on esophageal cancer cells.Up-regulation of GAS5 significantly promoted SOX6 protein level in esophageal cancer cell lines(P < 0.05),and co-transfection of GAS5 and miR-21 significantly reversed SOX6 protein(P<0.05).Conclusion: The expression of GAS5 was down-regulated in esophageal cancer tissues and cell lines,which was related to TNM stage.Overexpression of GAS5 inhibited the proliferation,invasion and migration of esophageal cancer cells,induced apoptosis,and plays an anti-cancer role in the evolution of esophageal cancer.GAS5 may inhibit the development of esophageal cancer by negatively regulating the expression of miR-21.GAS5 is involved in the occurrence and development of esophageal cancer by regulating the signal axis of miR-21/SOX6.