LncRNA TP73-AS1 Promotes Pancreatic Cancer Growth And Metastasis Through MiRNA-128/GOLM1 Axis

ObjectiveTo detect the expression level of lnc RNA TP73-AS1 in pancreatic cancer tissues and cell lines,and analyze its correlation with clinicopathological data.Further to explore the role and related mechanisms of TP73-AS1 in the development of pancreatic cancer.Materials and MethodsFirstly,the expression of TP73-AS1 in human pancreatic duct epithelial cell line(H6C7)and 5 other pancreatic cancer cell lines was detected,and then its expression level in 116 cases of pancreatic cancer tissues and corresponding adjacent tissues was detected.The in vitro study used CCK-8 experiment and clone formation experiment to detect the changes in cell proliferation after TP73-AS1 silencing;flow cytometry analyzed the changes in apoptosis of pancreatic cancer cells after down-regulation of TP73-AS1;the Transwell experiment was used to analyze the changes in the invasion ability of cells after knockdown of TP73-AS1.Using bioinformatics,it was found that TP73-AS1 acts as a competitive endogenous RNA(ce RNA)of miRNA-128-3p to regulate the expression of GOLM1.In vivo nude mouse tumor formation experiments evaluate the important role of TP73-AS1 in tumor growth and metastasis,and detect the expression of proliferation markers(Ki-67 staining)in xenograft tumor tissues.Results1.TP73-AS1 is significantly upregulated in pancreatic cancer tissues and cellsReal-time quantitative PCR showed that compared with H6C7 cells,TP73-AS1 was significantly up-regulated in pancreatic cancer cell lines and tissues.Increased TP73-AS1 expression was significantly correlated with tumor size,vessel infiltration,TNM staging and shorter overall survival.2.In vitro experiments to explore the biological function of TP73-AS1CCK-8 assay and clone formation experiment showed that knockdown of the expression of endogenous TP73-AS1 inhibited the proliferation and invasion of pancreatic cancer cells;flow cytometry analysis showed that the level of apoptosis increased after TP73-AS1 silencing.3.TP73-AS1 positively regulates GOLM1 by regulating the expression of miR-128-3pBioinformatics and dual-luciferase reporter analysis confirmed that TP73-AS1 and GOLM1 could be used as target genes of miR-128-3p,and rescue experiments elaborated that TP73-AS1 acted as a ce RNA of miR-128-3p to regulate the expression of GOLM1 to promote the progress of pancreatic cancer.4.Knockdown of TP73-AS1 could reduce tumorigenicity and metastasis in miceIn vivo experiments data showed that TP73-AS1 silencing could significantly reduce tumor growth volume.In addition,immunohistochemical staining indicated that the expression of Ki-67 was significantly lower in TP73-AS1 silencing group than that in the SCR group.And silencing TP73-AS1 significantly reduced lung metastasis.Conclusions1.Lnc RNA TP73-AS1 was significantly up-regulated in pancreatic cancer tissues and functioned as a proto-oncogene;2.Knockdown of TP73-AS1 could effectively inhibit the proliferation and invasion of pancreatic cancer cells by affecting cell apoptosis;3.Molecular mechanism showed that TP73-AS1 regulates the expression of GOLM1 by acting as a ce RNA of miR-128-3p to promote the progression of pancreatic cancer.