Ginsenoside RG3 inhibits the proliferation, migration, invasion of human osteosarcoma 143B cells and promotes apoptosis
10.3781/j.issn.1000-7431.2020.11.848
- Author:
Xiaohan MAO
1
Author Information
1. Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Cell movement;
Ginsenoside;
Osteosarcoma;
Proliferation
- From:
Tumor
2020;40(4):233-244
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effects of ginsenoside Rg3 on the proliferation, apoptosis, migration and invasion of osteosarcoma 143B cells and its possible mechanisms. Methods: Osteosarcoma 143B cells were treated with gradient concentrations of Rg3 (40, 50, 60, 70 and 80 µmol/L) for 24, 48 and 72 h, respectively, comparing with a control group of 143B cells treated with DMSO. The effects of Rg3 on the viability and proliferation of 143B cells were measured by MTT assay and colony-forming assay, respectively. Then the value of half-maximal inhibitory concentration (IC50) were calculated in order to screen the optimal drug concentrations and its action time. The apoptosis of 143B cells was measured by Hoechst 33258 staining and its cell cycle distribution was detected by FCM method. The migration and invasion abilities of 143B cells were detected by scratch wound healing assay and Transwell assay, respectively. The expression level of proliferating cell nuclear antigen (PCNA) mRNA was detected by real-time fluorescent quantitative PCR. The Western blotting was used to detect the proliferation marker PCNA and apoptosis-related proteins Bcl-2, Bax, cleaved-Caspase 3, as well as the expression level of migration-and invasion-related proteins matrix metalloproteinase-2 (MMP-2), MMP-7. Finally, Luciferase Gene Reportor System was used to screen the signaling pathways regulated by Rg3. The expression levels of protein kinase B (PKB, also known as AKT), phospho-AKT (p-AKT), cAMP-response element binding protein (CREB), and p-CREB were measured by Western blotting. Results: Comparing with the control group, Rg3 in the concentration of 40, 50, 60, 70 and 80 µmol/L could cause the inhibition of the proliferation of 143B cells which had a positive relationship with the concentration and action time (P < 0.001), and the IC50 of Rg3 to 143B cells was (51.00±3.46µmol/L. Furthermore, Rg3 inhibited the expression of PCNA at mRNA and protein levels (P < 0.05). In addition, Rg3 could also promote the apoptosis of 143B and inhibit their migration and invasion abilities (all P < 0.05). The expression levels of apoptosis-related protein Bcl-2 in 143B cells treated with Rg3 was down-regulated, while the expression level of the proteins Bax and cleaved-Caspase3 were up-regulated remarkably (all P < 0.05). The expression level of migration-and invasion-related proteins MMP-2 and MMP-7 were also decreased (all P < 0.05). At the same time, the transcriptional activity of CREB was impaired, and the expression levels of p-AKT, p-CREB were decreased remarkably (both P < 0.05). Conclusion: Rg3 can inhibit the proliferative, imaginational and invasive of osteosarcoma 143B cells, and also induced its apoptosis, which may be related with the depression of the activation of CREB signaling pathway.
