Expression Of MicroRNA-630 In Pancreatic Cancer Stem Cells And The Effect On Its Function

Background:Pancreatic cancer is known as the "king of cancers",and its prognosis is extremely poor,with insidious symptoms and difficult to diagnose and treat.(Micro RNAs(miRNAs)are a class of small non-coding RNA molecules that play a key role in the regulation of genetic expression,gastric cancer and esophageal cancer cells,however,its diagnostic and prognostic value in pancreatic cancer has not been found yet.Therefore,based on the current study,we propose to investigate the expression and functional effects of miR-630 in pancreatic cancer stem cells(PCSCs).Objective:To detect the expression of miR-630 in pancreatic cancer stem cells and to explore the effect of miR-630 on the stem cell-like properties of PCSCs.Methods:1.In vitro culture of pancreatic cancer cell line PANC-1 and normal pancreatic cells HPDE.2.Flow cytometry sorted and screened out PCSCs.3.q RT-PCR was performed to detect the expression of miR-630 in PCSCs.4.Using cell transfection,the cells were divided into the following groups according to transfection: mimic-NC group,miR-630 mimic group,inhibitor-NC group,miR-630 inhibitor group,and the effect of miR-630 on the self-renewal and maintenance of PCSCs was further analyzed by EDU staining assay,clone formation,and cell sphere-forming assay.5.A mouse subcutaneous xenograft tumor model was constructed to assess the tumorigenic ability of PCSCs with different treatments.They were divided into agomir-NC group,miR-630-agomir group,antagomir-NC group and miR-630-antagomir group,and the volume and weight of subcutaneous tumors were measured,and the gene expression in the tumors was detected by q RT-PCR.Results:1.miR-630 expression was significantly lower in PANC-1 cells compared to normal pancreatic cells HPDE(p < 0.05),while miR-630 expression was more significantly decreased in PCSCs cells(p < 0.05).2.miR-630 mimic group had significantly lower Ed U positive expression compared to mimic-NC group(p < 0.05),which indicated that cell proliferation was inhibited and low expression of miR-630 led to the opposite trend.Clone formation,cell sphere-forming assays showed that clone number and sphere-forming ability were significantly lower in the miR-630 mimic group,and the miR-630-inhibitor group led to the opposite trend(p < 0.05).In addition,the expression of Nanog and Oct4 was found to be significantly reduced in the miR-630 mimic group,while it was increased in the miR-630-inhibitor group(p < 0.05).3.The volume and weight of subcutaneous tumors were significantly decreased after miR-630 agomir injection,while miR-630 antagomir was increased(p < 0.05).Assays of miR-630 expression in tumors also showed that miR-630 agomir increased miR-630 expression,while miR-630 antagomir decreased miR-630 expression.Conclusion:1.miR-630 expression was reduced in PCSCs.2.miR-630 overexpression inhibited the proliferation and stem cell maintenance ability of PCSCs.3.miR-630 overexpression inhibited the growth of tumors in vivo.4.miR-630 can effectively inhibit the occurrence and development of pancreatic cancer and is expected to be a new target for pancreatic cancer gene therapy.