Cucurbitacin B inhibits proliferation and invasion of osteosarcoma 143b cells and promotes apoptosis / 肿瘤

ABSTRACT

Objective: To investigate the effects of cucurbitacin B (CuB) on the proliferation, invasion and apoptosis of osteosarcoma 143B cells, to explore the related molecular mechanisms. Methods: 143B cells were treated with different concentrations of CuB. The proliferation ability of 143B cells was detected by crystal violet staining and colony formation assay. The invasion ability of 143B cells was detected by Transwell chamber assay. The apoptosis rate and cell cycle distribution were detected by flow cytometry (FCM) method. The expression levels of epithelial-mesenchymal transition (EMT)-related markers including Vimentin, Snail and matrix metalloproteinase-9 (MMP-9) mRNAs and proteins were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The expression levels of apoptosis-related proteins [Bad, cleaved-caspase 3, cleaved-polyadenosine diphosphate ribose polymerase (PARP) and cyclin B1] and protein kinase B (PKB, Akt) signal pathwayrelated proteins [Akt, phospho-Akt (p-Akt), phosphatase and tensin homolog deleted on chromosome ten (PTEN) and phospho-PTEN (p-PTEN)] were detected by Western blotting. Results: Compared with the untreated control group, the proliferation and invasion of 143B cells treated with different concentrations of CuB were significantly inhibited (all P < 0.05). After treatment with 30 or 40 nmol/L CuB, the apoptosis rate of 143B cells was increased (both P < 0.05), the cell cycle was blocked in G2/M phase (both P < 0.01). The expression levels of MMP-9, Vimentin and Snail mRNAs and proteins were remarkably down-regulated (all P < 0.05) in 143B cells treated with CuB. The expression levels of apoptosis-related Bad, cleavedcaspase 3 and cleaved-PARP proteins were remarkably up-regulated (all P < 0.05) in 143B cells treated with CuB, the expression of cell cycle-related cyclin B1 protein was down-regulated (all P < 0.01) in 143B cells treated with CuB. At the same time, CuB reduced the expression level of p-Akt protein and increased the expression level of p-PTEN protein (both P < 0.05). Conclusion: CuB can inhibit the proliferation and invasion of osteosarcoma 143B cells, and promote apoptosis. These effects may be related to Akt pathway impediment and EMT processing attenuation.