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miR-502-3p regulates the proliferation and apoptosis of colorectal cancer stem cells by targeting GTPBP2 gene
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Abstract:
Objective: To investigate the molecular mechanism by which miR-502-3p regulates the proliferation, cell cycle and apoptosis of colorectal cancer stem cells (CCSCs) by targeting GTP binding protein 2 (GTPBP2) gene. Methods: The immunomagnetic bead sorting technique was used to sort CCSCs (CD133+CD44+ cells and CD133-CD44- cells) from colorectal cancer HCT116 cells, and the expression level of miR-502-3p in the sorted cells was detected by qPCR. CD133+CD44+ cells were divided into different groups according to different transfections using liposome method, namely miR-NC group, miR-502-3p group,si-miR-NC group, si-miR-502-3p group, miR-502-3p+vector group and miR-502-3p+GTPBP2 group. The mRNA expression levels of miR-502-3p and GTPBP2 in cells were detected using qPCR method. The proliferation rate, cell cycle and apoptosis rate were detected by MTT assay and flow cytometry, and the protein expression levels of Ki67, CDK1, Bcl2, BAX and GTPBP2 were detected by WB. Dual-luciferase reporter gene assay adopted to verify the targeting relationship between miR-502-3p and GTPBP2 gene. Results: The expression level of miR-502-3p in CD133+CD44+ cells was significantly lower than that in CD133-CD44- cells (P<0.01). Compared with the miR-NC group, the cell proliferation rate and the proportion of S phase cells were significantly reduced (all P<0.01), the apoptosis rate and proportion of cells in G0/G1 phase were significantly increased (all P<0.01), the protein expression of Ki67, CDK1 and Bcl2 was significantly down-regulated (all P<0.01), and BAX protein expression was significantly up-regulated (P<0.01) in the miR-502-3p group. miR-502-3p targetedly regulated the expression of GTPBP2. Overexpression of GTPBP2 could reverse the effects of up-regulation of miR-502-3p on the proliferation, cell cycle and apoptosis of CCSCs. Conclusion: Up-regulating the expression of miR-502-3p can inhibits the proliferation, arrests the cell cycle, and induces the apoptosis of CCSCs. The mechanism may be related to the overexpression of GTPBP2 gene.
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