| Objective:To investigate the expression of ABHD11-AS1 in four different pancreatic cancer cells(PANC-1,SW1990,Pa Tu8988 and Bx PC-3).This study was to explore pancreatic cancer cell proliferation,migration,invasion process of long non-coding RNA ABHD11-AS1’s functions,and analysis on the role of long non-coding RNA ABHD11-AS1 in pancreatic cancer cells epithelial mesenchymal transition(EMT)process,and the possible mechanism of this effect.Methods:1.QRT-PCR was used to detect the expression levels of ABHD11-AS1 in four pancreatic cancer cells(PANC-1,SW1990,Pa Tu8988 and Bx PC-3).2.Purchase sh-ABHD11-AS1 and sh EGFP interfering plasmids and overexpressed plasmids of pc DNA3.1-ABHD11-AS1 and pc DNA3.1.Two types of pancreatic cancer cells with relatively high expression of long non-coding RNA ABHD11-AS1 were transfected with shABHD11-AS1 plasmid(Pa Tu8988 and SW1990),and two types of pancreatic cancer cells with relatively low expression of long non-coding RNA ABHD11-AS1 were transfected with pc DNA3.1-ABHD11-AS1plasmid(PANC-1 and Bx PC-3).The transfection efficiency of shABHD11-AS1 plasmid and pc DNA3.1-ABHD11-AS1 plasmid in pancreatic cancer cells was determined.3.The effect of long non-coding RNA ABHD11-AS1 on the proliferation and clonal formation of pancreatic cancer cells was detected by cell proliferation assay and cloning assay.4.The effects of up-regulation and down-regulation of long non-coding RNA ABHD11-AS1 on the migration ability of pancreatic cancer cells were verified by Transwell migration test and wound healing assay.5.Transwell invasion assay was used to verify the influence of up-regulation and down-regulation of long non-coding RNAABHD11-AS1 on the invasion ability of pancreatic cancer cells.6.Western blot was used to detect the effect of long non-coding RNA ABHD11-AS1 on the protein level of the indicators related to EMT.7.Western blot was used to detect the relation of ABHD11-AS1 and proteins in the Hippo–YAPpathway.Results:1.ABHD11-AS1 expression was relatively high in Pa Tu8988 and SW1990 cell lines,but relatively low in PANC-1 and Bx PC-3 cell lines.2.In PaTu8988 and SW1990 cells with relatively high expression of ABHD11-AS1,plasmids containing sh-ABHD11-AS1 and sh-EGFP were infected,and q RT-PCR results showed that sh-ABHD11-AS1 had strong interference efficiency.At the same time,the expression levels of ABHD11-AS1 were relatively low in PANC-1 and Bx PC-3,which were transfected with the overexpression plasmid pc DNA3.1-ABHD11-AS1 and q RT-PCR detection showed that pc DNA3.1-ABHD11-AS1 had a strong overexpression effect.3.After interfering with ABHD11-AS1 in Pa Tu8988 and SW1990 cells,the ability of proliferation and clone formation of pancreatic cancer cells was weakened.Conversely,ABHD11-AS1 overexpression in PANC-1 and Bx PC-3 cells enhanced the proliferation and clonal formation ability of pancreatic cancer cells.4.After down-regulating ABHD11-AS1,the migration and invasion ability of Pa Tu8988 and SW1990 cells decreased,while the migration and invasion ability of PANC-1 and Bx PC-3cells increased after up-regulating ABHD11-AS1.5.After down-regulation of ABHD11-AS1,the expression level of epithelial marker protein in Pa Tu8988 and SW1990 cells increased,but the expression level about mesenchymal marker protein decreased.After upregulation of ABHD11-AS1,the expression level of epithelial marker protein in PANC-1 and Bx PC-3 cells decreased,and the expression level of mesenchymal marker protein increased.6.After ABHD11-AS1 was down-regulated,the expression levels of Hippo-YAP pathwayrelated proteins p-Lats1 and p-YAP in Pa Tu8988 and SW1990 cells were increased,and the expression levels of Lats1,YAP had no difference.After up-regulation of ABHD11-AS1,expression levels of p-Lats1,p-YAP and p-MOB1 proteins related to Hippo-YAP pathway in PANC-1 and Bx PC-3 cells decreased and the expression of YAP was increased.Conclusions:1、ABHD11-AS1 expressions were different in four pancreatic cancer cells.2 、 ABHD11-AS1 promotes the proliferation,migration and invasion of pancreatic cancer cells.3、ABHD11-AS1 may regulate epithelial mesenchymal transformation of pancreatic cancer cells through Hippo-YAP pathway. |
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