A study on mechanism of anti-tumor natural product Garcinone E inhibiting autophagic flux in ovarian cancer cells

SUN Xuemei; LEI Shunmei; WANG Chunlai; DAI Yiping; WEI Dan; YIN Fuqiang; LIU Xia

doi:10.3969/j.issn.1674-3806.2025.05.09

Abstract：Objective To explore the effect of anti-tumor natural product Garcinone E on autophagy function of ovarian cancer cells and its mechanism.Methods Ovarian cancer cells HeyA8 and SKOV3 were selected for experiments.Ovarian cancer cells were intervened with different concentrations of Garcinone E.Cell viability was detected by CCK-8 assay,and autophagic vesicle formation was detected by monodansylcadaverine(MDC)staining assay.The HeyA8 cell line and SKOV3 cell line stably expressing stubRFP-sensGFP-LC3 lentivirus were constructed to detect the changes in cellular autophagic flux.Co-localization of lysosomes with sensGFP-LC3 in ovarian cancer cells was detected by using lysosomal staining assay.Western blot assay was applied to detect the expression levels of cell cycle-related proteins p21,cyclin-dependent kinase 4(CDK4),cyclin-dependent kinase 7(CDK7),protein kinase B(Akt),microtubule-associated protein 1-light chain 3B(LC3B)and autophagy-related protein p62 in ovarian cancer cells.Results The viability of ovarian cancer cells was significantly decreased(P＜0.05)after intervention with Garcinone E.The half maximal inhibitory concentration(IC50)of Garcinone E on HeyA8 cells and SKOV3 cells were 5.93 μmol/L and 8.14 μmol/L,respectively.The MDC staining assay revealed an increase in the number of autophagosomes in the ovarian cancer cells after intervention with Garcinone E.Fluorescence microscopy revealed that Garcinone E blocked the autophagic flux in HeyA8 cells and SKOV3 cells stably transfected with stubRFP-sensGFP-LC3 lentivirus.The results of lysosomal staining assay showed that Garcinone E blocked the binding of autophagosomes to lysosomes.The results of Western blot assay showed that the intervention with Garcinone E up-regulated the protein expression levels of LC3B,p62 and p21 in ovarian cancer cells(P＜0.05)and down-regulated the protein expression levels of Akt,CDK4 and CDK7(P＜0.05).Conclusion Garcinone E can inhibit the autophagic flux of ovarian cancer cells and block the cell cycle of ovarian cancer cells,which has an anti-ovarian cancer effect and the potential to serve as a lead compound in the development of chemotherapeutic drugs for ovarian cancer.