Clinical Significance Of PRKAB1 In Pancreatic Cancer And Its Role In Development And Progression

Background and purpose:Pancreatic cancer is a kind of aggressive tumor which associated with high rates of mortality,which survival rate remains low over the past few decades with no significant change even if through surgical resection,radiation and chemotherapy,which 5-year survival rate as low as less than 20%.AMP activated protein kinase(AMPK)is an important energy sensing kinase,many reports suggested that AMPK may be a potential therapeutic target for the treatment of cancer;AMPK β 1 regulatory subunit encoded by the PRKAB1,which may be a positive regulator of AMPK activity,but the role of it that involved in the regulation of the process of pancreatic carcinogenesis and development is not clear.This research reveals the effect of PRKAB1 on the physiological activities of pancreatic cancer cells,elucidates the association between the expression level of PRKAB1 in the pathological features and clinical prognosis of pancreatic cancer,and offers novel ideas for the diagnosis and treatment of pancreatic cancer.Methods:1.Bioinformatics analysisIn this paper,the expression level and cellular localization of PRKAB1 in various tumors,including pancreatic cancer,and the relationship between its mutation site,frequency,and manner and the infiltration of various immune cells in the body were analyzed through the GEPIA 2.0 database,HPA database,c Bio Portal database,and TIMER 2.0 database.2.Relevant experimentalWestern blot analysis of PRKAB1 expression on pancreatic normal cells and pancreatic cancer cells.Evaluated PRKAB1 expression by immunohistochemistry(IHC)staining from 75 pancreatic cancer and 16 normal pancreatic tissue specimens.PRKAB1 expression and clinical information are summarized in for K-M survival estimations and correlation study.To examine the role of PRKAB1 in pancreatic cancer cell proliferation,migration and invasion,CCK-8,cell scratch and Transwell cell invasion assays were performed by constructing stable PRKAB1 knock-down and negative control cell lines.Result:1.GEPIA 2.0 analyzed that PRKAB1 highly expressed in diffuse large B cell lymphoma(DLBC),glioblastoma(GBM),thymoma(THYM)and pancreatic adenocarcinoma(PAAD);The expression of PRKAB1 was significantly different between pancreatic cancer and normal pancreatic tissues(P<0.05).2.HPA database analyzed PRKAB1 was mainly expressed in the cytoplasm,with a certain degree of expression in the nucleus.In addition,PRKAB1 was significantly highly expressed in pancreatic cancer tissues from the online IHC outcomes of pancreatic cancer tissues provided by HPA.3.The c Bio Portal database indicated that the mutation of PRKAB1 in 175 pancreatic cancer patients mainly comprised missense mutations,with a probability of0.6%.In addition,its primary mutation site is at the region of AMPK β 1 protein.4.To explore the relationship between PRKAB1 and the infiltration of various immune cells in the immune microenvironment,the database of TIMER 2.0 showed that PRKAB1 was positively correlated with the infiltration of CD4 +,Tregs and neutrophils(P < 0.05)and negatively regulated with the infiltration of B cells,NK cells and monocytes(P < 0.05).5.WB indicated that PRKAB1 expression in pancreatic cancer cell lines PANC-1and CFPAC-1 was significantly higher(P < 0.05)than normal pancreatic cell line h TERT-HPNE.6.The correlation between the high/low expression of PRKAB1 and the clinicopathological characteristics of pancreatic cancer patients was analyzed according to the results of IHC.The results showed that PRKAB1 was significantly related to the degree of tumor differentiation(P=0.013)and serum CEA(P=0.002).K-M survival curve analysis shows that patients with high expression of PRKAB1 often have poorer prognosis(P<0.05).7.CCK-8,cell scratch and Transwell cell invasion assays were performed in PANC-1,CFPAC-1 with PRKAB1 knock-down and negative control cell lines to evaluate the effect of PRKAB1 on pancreatic cancer cell proliferation,migration and invasion.Knocking down PRKAB1 inhibited pancreatic cancer proliferation,migration and invasion in pancreatic cancer cell lines(P<0.05).Conclusions:In this study,the gene of PRKAB1 may contribute to the development and progression of pancreatic cancer,which expression highly correlated to the patient pathology information especially in differentiation grape and serum CEA level that the higher PRKAB1 expression means worse prognosis.Moreover,PRKAB1 promotes pancreatic cancer proliferation,migration and invasion,which may be a novel biomarker to prognosis evaluation and therapeutic target.