| Objective: Pancreatic adenocarcinoma(PAAD)is one of the most malignant tumors of the digestive system and is the fourth leading cause of cancer death.Although some risk factors for PAAD had been identified,such as obesity and diabetes,the specific etiology is still unclear.Due to the hidden onset,high malignancy,and poor prognosis of PAAD,it is still essential to further understand the mechanism of the occurrence and development of PAAD and find potential diagnostic markers or therapeutic targets.MYEOV is a tumorassociated gene abnormally expressed in various tumors and is closely related to the occurrence and development of tumors.Rho GTPase activating protein 1(ARHGAP1)is a tumor-related gene abnormally expressed in various tumors.It is correlated with the expression of MYEOV in pancreatic cancer and is closely related to the occurrence and development of tumors.However,the biological function and specific mechanism of MYEOV and ARHGAP1 in PAAD are not fully understood,so further studies are needed.Methods: 1.UCSC Xena database was used to analyze the expression level of MYEOV in PAAD and normal tissues,and the ICGC database was used to analyze the relationship between MYEOV and the prognosis of PAAD patients2.The expression of MYEOV in PAAD cells was detected by quantitative fluorescence PCR.3.The overexpression plasmid and knockdown plasmid were designed and transfected into PAAD cell lines,respectively.The up-regulation and down-regulation experiments of MYEOV expression were performed,and the empty plasmid was used as the control group.4.Explore the biological function of MYEOV : upregulate or down-regulate the expression of MYEOV,and conduct cell physical function studies such as cell proliferation,migration,and invasion.5.To explore the mechanism of MYEOV: Western blot was used to detect the expression levels of matrix metalloproteinases MMP2 and MMP9.Western blot was used to detect the role of MYEOV in EMT(EMT).The expression levels of p-Smad2/3 and TGFβ-1 in the TGFβ/Smad signaling pathway were detected by Western blot.6.Explore the mechanism of MYEOV and ARHGAP1 : the correlation between MYEOV and ARHGAP1 was analyzed by reviewing relevant RNAseq and databases,and the correlation between them was detected by co-immunoprecipitation.7.Western blot was used to detect the expression of ARHGAP1 in PAAD cells.8.ARHGAP1 small interfering RNA was designed and transfected into PAAD cell lines respectively for the ARHGAP expression down-regulation experiment.The transfection of NControl-si RNA was set as the control group.9.Explore the biological function of ARHGAP1:The expression of ARHGAP1 was down-regulated to study the natural function of cells,such as cell proliferation,migration,invasion,and apoptosis.10.To explore the mechanism of ARHGAP1:Western blot was used to detect the expression levels of matrix metalloproteinases MMP2 and MMP9.Western blot was used to detect the role of ARHGAP1 in EMT(EMT).Results: 1.The expression of MYEOV was analyzed by the UCSC Xena database,and the results showed that the expression of MYEOV was significantly increased in PAAD samples;Kaplan-Meier analysis of the ICGC database showed that the higher the expression of MYEOV,the shorter the median survival time of PAAD patients.2.Among the five PAAD cell lines PaTu8988,SW1990,BXPC3,Mia PaCa-2 and PANC1,the expression of MYEOV was higher in Pa Tu8988 cells and lowest in Mia Pa Ca-2 cells.Therefore,the knockdown effect of MYEOV on PAAD was investigated in Pa Tu8988 cells,and the effect of overexpression of MYEOV on PAAD was investigated in Mia Pa Ca-2 cells.3.Cell function experiment showed that the knockdown of MYEOV could inhibit the proliferation,migration,and invasion of PAAD cells.The opposite trend was observed after the overexpression of MYEOV.4.Western blot results showed that the expression levels of matrix metalloproteinases MMP2 and MMP9 were significantly decreased after knocking down MYEOV.Western blot results showed that MYEOV knockdown increased the protein expression level of epithelial marker E-cadherin,and decreased the protein expression levels of mesenchymal markers N-cadherin and Vimentin.In contrast,overexpression of MYEOV showed the opposite trend.5.Western blot results showed that the expression levels of p-Smad2/3 and TGFβ-1 were significantly decreased after the knockdown of MYEOV,while the expression levels of p-Smad2/3 and TGFβ-1 were significantly increased after overexpression of MYEOV.6.Relevant literature reports and database analysis showed that MYEOV was related to the expression of ARHGAP1.Co-immunoprecipitation results showed that MYEOV could interact with ARHGAP1,and MYEOV affected the expression of ARHGAP1 protein.7.In Pa Tu8988,SW1990,Bx Pc 3,Mia Pa Ca-2,and PAN1 of five pancreatic adenocarcinoma cell lines,the protein expression of ARHGAP1 in SW1990,Bx Pc 3 cells were relatively high.Therefore,the effect of ARHGAP1 knockdown on pancreatic adenocarcinoma was investigated in SW1990 and BXPC3 cells.8.The proliferation,migration,and invasion ability of pancreatic adenocarcinoma cells were inhibited when knockdown ARHGAP1.9.Western blot results showed that the expression levels of matrix metalloproteinases MMP2 and MMP9 were significantly decreased after ARHGAP1 knockdown.However,the protein expressions of mesenchymal markers N-cadherin and Vimentin were downregulated.Conclusions:In conclusion,the expression of MYEOV was upregulated in PAAD tissues and cells,and the expression level was closely related to the survival of patients.MYEOV promoted the proliferation,migration,and invasion of PAAD cells,and MYEOV may activate the EMT of PAAD cells through the TGFβ/Smad signaling pathway.MYEOV was associated with the expression of ARHGAP1 in PAAD,and MYEOV could affect the expression level of ARHGAP1 protein in PAAD cells.ARHGAP1 was upregulated in PAAD tissues.ARHGAP1 promoted the proliferation,migration,and invasion of PAAD cells.These results suggested that MYEOV may interact with ARHGAP1 to affect the occurrence and development of PAAD and may be a potential biomarker or a new target for the treatment of PAAD patients.Figure[15]Table[30]Reference[47]... |
PDF Full Text Request