| BACKGROUNDPancreatic cancer(PC)is a highly malignant disease,and its molecular mechanism is unclear.A recent study demonstrated the upregulated expression of the long noncoding RNA(lncRNA)EPIC1 in cancer.However,no studies have investigated the relationship between Lnc-EPIC1 and PC.Therefore,elucidating the underlying mechanisms of Lnc-EPIC1 may be helpful in identifying novel diagnostic and therapeutic targets for PC.AIMTo investigate the expression of lnc-EPIC 1 in PC cell lines and PC tissues,the correlation between lnc-EPIC 1 and PC tumor size,TNM stage,and lymph node metastasis in clinical samples.The role of apoptosis and the signaling pathway of lnc-EPIC1 in PC.METHODSThe expression of Lnc-EPIC1 in 30 PC tissues and human PC cells was detected by qRT-PCR,and the correlation between Lnc-EPIC1 and clinicopathological features(Tumor size,TNM stage,lymph node metastasis)was analyzed.The expression of YAP 1 in tissues and cells was detected by Western blotting.The effect of YAP1 on the staging and prognosis of PC was determined by TCGA database analysis.Subsequently,after transfection of siRNA into PC cells,qRT-PCR technology to detect transfection efficiency and take pictures.We used the CCK-8 assay and colony formation assay to detect cell proliferation and cloning ability.In addition,we used flow cytometry to measure the apoptosis rate and cell cycle progression.CatRAPID graphic software was used to analyze UKB database data to predict the interaction tendency and binding site of lnc-EPIC 1 and YAP1.We verified the mechanism of action of lnc-EPIC 1 in PC by RNA immunoprecipitation.RESULTSThe qRT-PCR test showed that the expression of Lnc-EPIC 1 in PC cell lines(SW1990 cell line,HPAF? cell line)was significantly increased compared with normal pancreatic cell lines(HPDE cell lines)(P<0.001).Analysis of clinical medical records showed that the expression level of lnc-EPIC 1 was up-regulated in PC and was positively correlated with tumor size,TNM stage,and lymph node metastasis(P<0.01,P<0.01,P<0.01).Western blot results showed that compared with the normal pancreatic cell line(HPDE cell line),the expression of YAP1 was significantly increased in PC cell lines(SW1990 cell line,HPAF ? cell line).At the same time,the expression of YAP1 in PC tissues was also significantly higher than that in PC tissues.GEPIA's TCGA-PAAD data analysis showed that compared with normal pancreatic tissue,the expression of YAP 1 was higher in PC tissues(P<0.05).The overall survival rate of PC patients with high expression of YAP 1 was poor(P<0.01),and the tumor stage was higher(P<0.001).Lnc-EPIC1 and YAP1 are highly expressed in PC cells and tissues,which may be related to PC progression.Targeted siRNA(EPIC1 siRNA 1,EPIC1 siRNA2)can significantly inhibit the expression of lnc-EPIC 1(p<0.001,p<0.001).After low expression of lnc-EPIC 1,the cell proliferation ability of PC cell lines(SW1990 cell line,HPAF? cell line)was significantly reduced(p<0.001,p<0,001),and cell cloning function was significantly reduced(p<0.001,p<0.001),Apoptosis rate was significantly increased(p<0.001,p<0.001).In the SW1990 cell line,qRT-PCR results showed that after low expression of Lnc-EPIC1,the percentage of G0/1 cells increased significantly(p<0.001),G1/S cell cycle arrest,cyclin Cyclin Al,CDC20 and CDK4 mRNA.The expression levels were significantly reduced(P<0.001,P<0.001,P<0.001).Using bioinformatics to analyze the correlation between YAP1 and lnc-EPIC 1,cat RAPID software can identify the interaction area of YAP 1 and lnc-EPIC 1 with a high degree of confidence(interaction score=135,discriminative power=100%),meanwhile,the possible binding sites of YAP 1 and lnc-EPIC1 are located at 600BP and 1300BP.RIP results showed that there was a direct interaction between lnc-EPIC 1 and YAP 1(p<0.01).CONCLUSIONS1.Lnc-EPIC 1 is highly expressed in PC cells and PC tissues;2.The expression of lnc-EPIC 1 is related to tumor size,TNM stage and lymph node metastasis in PC;3.Downregulating the expression level of lnc-EPIC 1 reduces the tumorigenic function of PC cells and increases the apoptosis rate;4.Lnc-EPIC1 targets YAP1,regulates cyclins Cyclin A1,CDC20,and CDK4,and promotes the progression of PC. |
PDF Full Text Request