Objective To investigate the effects of fructose-2-kinase 6-phosphate/fructose-2,6-diphosphatase 4(PFKFB4)on the stemness of ovarian cancer cells and its mechanism.Methods Ovarian cancer SKOV3 and A2780 stable cell lines with PFKFB4 overexpression were constructed by lentivirus system.Real-time PCR and western blot analysis was performed to detect the expression of stemness-related transcription factors SOX2,OCT4 and NANOG.The sphere-forming ability of the cells was detected by sphere formation assay to determine the effect of PFKFB4 on the stemness of ovarian cancer cells.The expression of SOX2,OCT4 and NANOG and the sphere-forming ability of the cells were detected after treatment with rapamycin,an autophagy activator,to evaluate the effect of autophagy on the stemness of ovarian cancer cells.The expression of autophagy-related proteins LC3B-Ⅱ and p62 was detected by western blot analysis to determine the effect of PFKFB4 on autophagy of ovarian cancer cells.The correlation between the expression of PFKFB4 and BNIP3 in ovarian cancer was analyzed by bioinformatics.Real-time PCR and western blot analysis was used to detect the effect of PFKFB4 on the expression of BNIP3.Results Overexpression of PFKFB4 promoted the expression of SOX2,OCT4 and NANOG and the sphere-forming ability of ovarian cancer cells.Rapamycin increased the expression of SOX2,OCT4 and NANOG and the sphere-forming ability of ovarian cancer cells.Overexpression of PFKFB4 increased the autophagy level of ovarian cells,including increasing the expression of LC3B-Ⅱ protein and decreasing the expression of p62 protein.Bioinformatics analysis showed that PFKFB4 and BNIP3 expression were highly positively correlated in ovarian cancer.Overexpression of PFKFB4 increased the mRNA and protein levels of BNIP3.Conclusion PFKFB4 may enhance the stemness of ovarian cancer cells by activating BNIP3-mediated autophagy.
维普
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