ABSTRACT
Objective: To investigate the effect of silencing sirtuin 3 (Sirt3) on the apoptosis of human ovarian cancer SKOV3 cells induced by resveratrol (Res), and to explore its mechanism of promoting apoptosis. Methods: The human ovarian cancer SKOC3 cells were cultured with different concentrations 0, 2. 5, 5. 0, 10.0, 20.0, 40.0 and 80.0 mg · L-1) of Res for 24 h. The survival rate of cells was measured by MTT assay. The SKOV3 cells were randomly divided into control group, Sirt3 inhibitory 3-1H-1, 2, 3-triazol-4-yl) pyridine 3-TYP group, Res group and 3-TYP+Res group. After 24 h of culture, the inhibitory rates of proliferation of the cells in various groups were detected by MTT assay; the nuclei were stained with Hoechst 33342, and the morphorgy nucleus was observed by laser confocal microscope; reactive oxygen species (ROS) probe was used to detect the intracellular ROS levels; Western blotting method was used to detect the expression levels of Sirt3, Bax, Bcl-2 and cleaved caspase-3 proteins in the cells in various groups. Results: The results of MTT assay showed that the survival rates of SKOV3 cells were significantly decreased with the increase of concentration of Res, and the median inhibitory concentration (IC50) was 42. 73 mg · L-1. Compared with control group, the inhibitory rates of proliferation of cells in Res group and 3-TYP+Res group were significantly decreased (P0.05); the protein expression levels of Sirt3 and Bcl-2 proteins in Res group were significantly decreased (P< 0.05), and the expression levels of Bax and cleaved caspase-3 proteins were significantly increased (P<0.05). Compared with Res group, the expression levels of Bax and cleaved caspase-3 proteins in 3-TYP + Res group were significantly increased (P<0.05), and the expression levels of Bcl-2 and Sirt3 proteins in 3-TYP+Res group were significantly decreased (P<0.05). Conclusion: Res can induce the apoptosis of SKOV3 cells, and the inhibition of Sirt3 expression by 3-TYP can enhance the effect of Res.
ABSTRACT
Aim To investigate the effect of Rhein derivative 4a containing amide structure on migration and invasion in ovarian cancer SKOV3 cells and its possible mechanism. Methods Ovarian cancer SK0V3 cells were used as target cells. Molecular docking and West-em blot were used to detect the regulatory effect of derivative 4a on Racl protein. CCK8, HE staining, Scratch and Transwell assay were used to detect the effects of derivative 4a on the proliferation, morphology , migration and invasion of SK0V3 cells, respectively. Western blot was employed to determine the expression of matrix metalloproteinases and EMT-related proteins. Results Derivative 4a could effectively bind to Racl protein, and the binding energy was-29. 10 kcal • mol"1, which was significantly lower than that of Rhein; it also could down-regulate the expression of Racl protein in SK0V3 cells. Derivative 4a could significantly inhibit the proliferation, invasion and migra tion of SKOV3 cells, and induce a large amount of cellular vacuolation; derivative 4a could also down-regu-\ late the expression of MMP-2 and MMP-9, up-regulate the expression of EMT epithelial marker protein E-ca-derin but down-regulate the expression of vimentin and j3-cantenin. Conclusions Derivative 4 a can inhibit the proliferation, migration and invasion of ovarian cancer SK0V3 cells. The mechanism may relate to its targeted regulation of Racl, thereby inhibiting the secretion of matrix metalloproteinases, up-regulating the expression of key molecule E-caderin and down-regula-ting the expression of Vimentin and (3-cantenin in EMT i process.•.
ABSTRACT
Objective:To investigate the effect of berberine on the proliferation and apoptosis of human ovarian cancer cell (SKOV3). Methods:Cell proliferation was detected by MTT method. The cell apoptosis was detected by FCM Annexin V/PI double staining and transmission electron microscopy. The methylation status of hMLH1 gene promoter CpG island was analyzed by methylation specific PCR. The expression of Bcl-2, Bax, Survivin and hMLH1 gene mRNA were detected by real-time fluorescent quantitative RT-PCR. Results:The berberine could significantly inhibit the proliferation of ovarian cancer SKOV3 cells(P<0. 05) in dose-and time-de-pendent manner. When combined with cisplatin, berberine showed synergistic anticancer effects. Berberine could induce SKOV3 cells apoptosis significantly, it might lower the expression of Bcl-2 and Survivin gene and enhance the expression of Bax gene. In addition, berberine could restore the hMLH1 promoter methylation status and increase the expression of hMLH1 mRNA. Conclusion:Berberine can inhibit the proliferation of ovarian cancer cells and induce apoptosis, which show that the synergistic enhancement anticancer effects with cisplatin.
ABSTRACT
Objective:To investigate the effect of berberine on the proliferation and apoptosis of human ovarian cancer cell (SKOV3). Methods:Cell proliferation was detected by MTT method. The cell apoptosis was detected by FCM Annexin V/PI double staining and transmission electron microscopy. The methylation status of hMLH1 gene promoter CpG island was analyzed by methylation specific PCR. The expression of Bcl-2, Bax, Survivin and hMLH1 gene mRNA were detected by real-time fluorescent quantitative RT-PCR. Results:The berberine could significantly inhibit the proliferation of ovarian cancer SKOV3 cells(P<0. 05) in dose-and time-de-pendent manner. When combined with cisplatin, berberine showed synergistic anticancer effects. Berberine could induce SKOV3 cells apoptosis significantly, it might lower the expression of Bcl-2 and Survivin gene and enhance the expression of Bax gene. In addition, berberine could restore the hMLH1 promoter methylation status and increase the expression of hMLH1 mRNA. Conclusion:Berberine can inhibit the proliferation of ovarian cancer cells and induce apoptosis, which show that the synergistic enhancement anticancer effects with cisplatin.