Molecular Mechanism of circVRK1 Regulating the Proliferation and Apoptosis of Acute Lymphoblastic Leukemia KOCL44 Cells by Targeting miR-4428
Objective To elucidate the role of circVRK1 and its interaction with miR-4428 in regulating proliferation and apoptosis in acute lymphoblastic leukemia(ALL)cells.Methods KOCL44 ALL cells were cultured in vitro,and experimental groups included pcDNA,pcDNA-circVRK1,anti-miR-NC,anti-miR-4428,si-NC,si-circVRK1,pcDNA-circVRK1+miR-NC,and pcDNA-circVRK1+miR-4428.The expression levels of circVRK1 and miR-4428 were detected using qRT-PCR.CCK-8 assays and flow cytometry were used to assess cell proliferation and apoptosis,respectively.The dual luciferase reporter assays were employed to investigate the interaction between circVRK1 and miR-4428,with groups categorized as WT-circVRK1+miR-NC,WT-circVRK1+miR-4428,MUT-circVRK1+miR-NC,and MUT-circVRK1+miR-4428.Western blotting was utilized to detect the expression levels of Ki-67,cleaved caspase-3,and cleaved caspase-9 proteins.Results Compared to the pcDNA group,circVRK1 expression was up-regulated in the pcDNA-circVRK1 group(P＜0.05).Compared to transfection with pcDNA or anti-miR-NC,transfection with pcDNA-circVRK1 or anti-miR-4428 led to decreased cell viability and Ki-67 protein levels in KOCL44 cells(P＜0.05),and increased apoptosis rates and levels of cleaved caspase-3 and cleaved caspase-9(P＜0.05).circVRK1 was found to negatively regulate miR-4428 expression,with this effect observed only in the WT-circVRK1 group.miR-4428 levels were lower in the pcDNA-circVRK1 group compared to the pcDNA group(P＜0.05)and higher in the si-circVRK1 group compared to the si-NC group(P＜0.05).Co-transfection with pcDNA-circVRK1+miR-4428 resulted in increased cell viability(P＜0.05)and Ki-67 expression(P＜0.05),and decreased apoptosis rates and levels of cleaved caspase-3 and cleaved caspase-9(P＜0.05)compared to co-transfection with pcDNA-circVRK1+miR-NC.Conclusion Overexpression of circVRK1 reduces the proliferation ability of acute ALL cells and induces cell apoptosis by downregulating miR-4428 expression.
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