| Background:Recently,increasing studies has revealed that aberrant expression of long non-coding RNAs(lncRNAs)in a variety of cancers involved in cancer cell proliferation,apoptosis,invasion and metastasis.Our previous studys have screened aberrant expression of lncRNA CRNDE in normal pancreatic tissues and pancreatic cancer tissues by using lncRNA Microarray,however,its role in pancreatic cancer is still unclear.Objective: This study was aimed to discuss the role of CRNDE in pancreatic cancer on proliferation,apoptosis and invasion,and to explore the role of Hippo-YAP pathways mediated by CRNDE in malignant transformation of pancreatic cancer progression.Methods:1.We detected the expression of CRNDE in 20 cases of pancreatic carcinoma tissues,adjacent normal tissues and 3 types of pancreatic cancer cells(AsPC-1,BxPC-1,PANC-3)by RT-PCR assay,and using flow cytometry and Transwell assay to determine the effects of down-regulation of CRNDE on pancreatic cancer cell proliferation,apoptosis and migration.2.In this study,we investigated the expression levels of YAP1 and LAST1 protein in pancreatic cancer tissue by IHC and analysed the relationship among CRNDE,YAP1,LAST1.We explored the expression of YAP1 and LAST1 when the expression of CRNDE in pancreatic cancer cells was down-regulated by Western blot and elucidated the role of CRNDE mediating Hippo-YAP pathway in the mechanism of cell malignant transformation in pancreatic cancer.Results:1.In this study,we detected the expression of CRNDE in pancreatic cancer tissues and adjacent normal tissues of 20 cases.The results confirmed that the expression levels of CRNDE in pancreatic cancer tissues was significantly higher than that inadjacent normal tissues by RT-PCR(P<0.05).The expression of CRNDE also varied in AsPC-1,PANC-1,BxPC-3.The AsPC-1 cell which was expressed highest were applied to the experiment of loss-of-function.Flow cytometry detection found that G1 phase cells decreased significantly,while G2/M phase cell increased obviously(P<0.05)in the experimental group,and the rate of cell apoptosis increased significantly compared with that in the control group(P<0.05).Transwell experiment confirmed that the ability of invasion of cancer cells in the experimental group was significantly inhibited(P<0.05).2.The level of LATS1 decreased significantly and the level of YAP1 expression increased obviously in pancreatic cancer tissues compared with that in adjacent normal tissues.Besides,linear regression analysis showed that CRNDE was related to the LAST1 and YAP1.There was a negative correlation between CRNDE and LAST1,meanwhile there was a positive correlation existed between CRNDE and YAP1.LATS1 protein expression levels increased and YAP1 protein expression levels decreased when CRNDE expression was down-regulated in pancreatic cancer AsPC-1 cells by Western blot.Conclusions:1.CRNDE was highly expressed in pancreatic cancer,and it promoted the growth of tumor cells,inhibited cell apoptosis and enhanced tumour invasiveness.This results confirmed that CRNDE is essential for the development of pancreatic cancer.2.Down-regulation of CRNDE expression in pancreatic cancer cells could significantly increase LATS1 expression and inhibit YAP1 expression.Furthermore,the results showed that CRNDE could mediate Hippo-YAP signaling pathway which plays an important role in the development of pancreatic cancer. |
PDF Full Text Request