Effect Of PiRNA MW557525 On Proliferation,Apoptosis,Migration And Invasion Of Piwil2-iCSCs And Wilms Tumor Cells G401

Objective : To investigate the effect of the differential piRNA MW557525 derived from Piwil2-induced cancer stem-like cells(Piwil2-iCSCs)on the biological behavior of Piwil2-iCSCs and Wilms tumor cells(G401).Methods:1 High-throughput sequencing(HTS)was used to screen differential piRNAs of Piwil2-iCSCs.Target genes were predicted by the miRanda algorithm and subjected to Gene Ontology(GO)and KEGG analysis.2 RT-qPCR was used to detect the expression of piRNA MW557525 and NOP56 in Piwil2-iCSCs and Wilms tumor cell line G401.3 Piwil2-iCSCs and G401 were transfected with piRNA MW557525 Mimics and Inhibitor to over-express and inhibit its expression.RT-qPCR was used to verify the expression of piRNA MW557525 in Piwil2-iCSCs and G401 cells after transfection.4 The proliferation of Piwil2-iCSCs and G401 cells after transfection were detected by CCK8,the apoptosis ability were detected by flow cytometry,and the healing assay and Transwell were used to detect cell migration and invasion capacity;the expression of BAX,BCL2,MMP2 and MMP9 were detected by Western blot.5 The binding of NOP56 and piRNA MW557525 was detected by dual luciferase experiment;the interaction between NOP56 and piRNA MW557525 were detected by Western blot and RT-qPCR.6 The expression of stem cell pluripotency-related markers CD24,CD133,SOX2 and KLF4 in Piwil2-iCSCs were detected by Western blot.Results:1 Via HTS,we screened out 204 differential piRNAs,and miRanda predicted 77 target genes.GO analysis showed that the biological processes(BPs)of these target genes were mainly involved in regulating the calcium concentration of cells and their molecular functions(MFs)were mainly involved in ATPase activity.2 RT-qPCR showed that levels of piRNA MW557525 were significantly upregulated in Piwil2-iCSCs than that in FB,while the expression of piRNA MW557525 in human Wilms tumor G401 cells decreased significantly compared with renal tubular epithelial cells HK2;NOP56 were highly expressed in Piwil2-iCSCs and G401;piRNA MW557525 did not bind to NOP56 directly,but regulated the expression of NOP56.3 PiRNA MW557525 promoted proliferation,migration,invasion and pluripotency and inhibited apoptosis,while NOP56 suppressed proliferation,migration,invasion and pluripotency and induced apoptosis,in Piwil2-iCSCs.4 PiRNA MW557525 obviously suppressed the proliferation,migration and invasion of G401 cells and promoted cell apoptosis;PiRNA MW557525 inhibited the expression of MMP2,MMP9 and BCL2,and promoted the expression of BAX.Conclusion:1 PiRNA MW557525 is highly expressed in Piwil2-iCSCs and low in G401;NOP56 is highly expressed in both Piwil2-iCSCs and Wilms tumor.PiRNA MW557525 may indirectly regulate the expression of NOP56.2 PiRNA MW557525 can promote the proliferation,migration,invasion and stem cell pluripotency of Piwil2-iCSCs and inhibit the apoptosis of Piwil2-iCSCs.NOP56 can promote the apoptosis and inhibit the proliferation,migration,invasion and the pluripotency of Piwil2-iCSCs.3 PiRNA MW557525 can inhibit the proliferation,migration and invasion and promote the apoptosis of G401 cells.