Linc-RoR Promotes Stemness Properties Via The Wnt/β-catenin Signaling Pathway In Pancreatic Cancer Cells

Pancreatic cancer is one of the most destructive and deadly digestive tract malignancies in the world.Due to the insidious onset of pancreatic cancer and the lack of early characteristic clinical manifestations,early diagnosis is difficult.Most patients with pancreatic cancer have been diagnosed in the advanced stage and have lost the best opportunity for surgical treatment.In addition,the therapeutic effect of chemoradiotherapy on pancreatic cancer is poor,and currently effective immunotherapy methods are scarce.Meanwhile,due to the unknown pathogenesis of pancreatic cancer,rapid progression and early metastases,resulting in a very poor prognosis,with a 5-year survival rate of less than 5%.Therefore,it is particularly important to explore the relevant molecular mechanisms of the development of pancreatic cancer and lay a foundation for further clinical research on gene targeted therapy for pancreatic cancer.Tumor stem cells(CSC)are a few undifferentiated cells in tumor cells that are in a state of resting division.They have the ability of self-renewal,as well as the unique plasticity,metabolism and enhanced chemoradiotherapy resistance.Studies have shown that in pancreatic cancer,pancreatic cancer stem cells(PCSC)drive the development and progression of the tumor and play an important role in tumor metastasis,recurrence and drug resistance.The highly malignant biological behavior of pancreatic cancer may be determined by pancreatic cancer stem cells in pancreatic cancer tissues.Long non-coding RNA(lncRNA)is a non-coding RNA molecule composed of more than 200 nucleotides that do not encode proteins.A number of studies have shown that lncRNA is a key regulatory factor in various cell biological behaviors such as embryo development and cell metabolism,and its abnormal expression is closely related to tumor proliferation,invasion and metastasis.Linc-RoR is a classic long non-coding RNA with a length of 2.6kb.It is an important target of the main transcription factors of stem cells,and is believed to play a promoting role in a variety of malignant tumors such as lung cancer,gastric cancer,endometrial cancer and so on.Objective: To explore the regulatory effect of linc-RoR on the stemness of pancreatic cancer cells and its related molecular biological mechanisms,and lay a foundation for further clinical research on gene targeted therapy for pancreatic cancer.Methods:1.The expression of linc-RoR in pancreatic cancer cell lines Pa Tu-8988,SW1990 and Bx PC-3 was detected by Real-time PCR.2.According to the expression level,pCDH-RoR plasmid was transfected into pancreatic cancer cells with low linc-RoR expression(Bx PC-3,SW1990),and sh-RoR plasmid was transfected into pancreatic cancer cells with relatively high linc-RoR expression(SW1990,Pa Tu-8988),and the transfection efficiency was detected.3.The regulation of linc-RoR on the stemness of pancreatic cancer cells was investigated by laboratory techniques such as Real-time PCR,Western blot and stem cells sphere formation.4.The expression of linc-RoR in pancreatic cancer cells was up-regulated or down-regulated,and the expression levels of proteins associated with the Wnt/β-catenin signaling pathway were detected by Western blot.Results:1.The results of Real-time PCR showed that linc-RoR was differentially expressed in three pancreatic cancer cells,among which the expression of linc-RoR in Pa Tu-8988 was the highest,the expression in SW1990 was relatively low,and the expression of linc-RoR in Bx PC-3 was the lowest.2.The expression of linc-RoR in pancreatic cancer cells was up-regulated in Bx PC-3and SW1990,and down-regulated in SW1990 and Pa Tu-8988.3.Linc-RoR is involved in regulating the stemness of pancreatic cancer cells:3.1 After up-regulating Linc-RoR,the protein level of stemness related genes detected by Western blot was significantly increased.On the contrary,after down-regulating Linc-RoR,the protein level of stemness related genes was significantly reduced.3.2 After up-regulating Linc-RoR,the m RNA level of stemness related genes detected by Real-time PCR was significantly increased.Conversely,down-regulation of Linc-RoR expression in pancreatic cancer cells significantly reduced the level of m RNA expression of stemness related genes.3.3 After up-regulating Linc-RoR,the sphere formation ability of pancreatic cancer cells was significantly higher than that of control cells.After down-regulating the expression of Linc-RoR,the sphere formation ability of pancreatic cancer cells was significantly decreased.4.Linc-RoR promotes the stemness of pancreatic cancer cells by regulating the Wnt/β-catenin signaling pathway:4.1 After up-regulating Linc-RoR,the expression levels of Wnt/β-catenin signaling pathway related proteins(p-GSK3 β,β-catenin,Cyclin D1,Met)were significantly increased,while the total GSK-3 β protein expression level was not significantly changed.4.2 After down-regulating Linc-RoR,the expression levels of Wnt/β-catenin signaling pathway related proteins(p-GSK3 β,β-catenin,Cyclin D1,Met)were significantly decreased,while the total GSK-3 β protein expression level was not significantly changed.Conclusion:1.Linc-RoR was differentially expressed in different pancreatic cancer cell lines.2.Linc-RoR is involved in regulating the stemness of pancreatic cancer cells.Up-regulating expression of Linc-RoR can promote the stemness of pancreatic cancer cells,while down-regulating expression of Linc-RoR can inhibit the stemness of pancreatic cancer cells.3.Linc-RoR promotes stemness in pancreatic cancer cells by regulating the Wnt/β-catenin signaling pathway.