Effect of silencing sirtuin 3 on resveratrol-induced apoptois of human ovarian cancer SKOV3 cels
10.13481/j.1671-587x.20200108
- Author:
Songyan LI
1
Author Information
1. Key Laboratory of Tumor Targeted Therapy and Transformation Medicine, School of Basic Medical Sciences, Jilin Medical University
- Publication Type:Journal Article
- Keywords:
3-(1H-1, 2, 3-triazol-4-yl) pyridine;
Apoptosis;
Human ovarian cancer SKOV3 cells;
Resveratrol;
Sirtuin 3
- From:
Journal of Jilin University(Medicine Edition)
2020;46(1):45-49
- CountryChina
- Language:Chinese
-
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 (P<0.05); compared with Res group, the inhibitory rate of proliferation of the SKOV3 cells in 3-TYP +Res group was significantly inereased (P<0.01). Compared with control group, the nucleus of SKOV3 cells in 3-TYP + Res group showed pyknosis, enhanced staining and more nuclear fragmentation. Compared with control group, the red fluorescence of the SKOV3 cells in 3-TYP group had no significant change, and the red fluorescence of the SKOV3 cells in Res group and 3-TYP+Res group was significantly decreased. The results of Western blotting method showed that compared with control group, the expression level of Sirt3 protein in the cells in 3-TYP group was significantly decreased (P<0.05), and the expression levels of Bcl-2, Bax and cleaved caspase-3 in the cells did not change significantly (P>0.05); 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.
