RESUMO
Objective:To explore the potential mechanism and feasibility of sonic hedgehog (SHH) signal pathway inhibitor NVP-LDE225 combined with radiotherapy in the treatment of melanoma.Methods:The Gli1 mRNA expression of the melanoma cells (Melan-A and SK-MEL-2) was detected by qPCR. After treatment with NVP-LDE225, GDC-0449 or combined with radiation for 24 hours, the number and activity of Melan-A and SK-MEL-2 were detected by cell counting and CCK-8 kit. The survival and proliferation ratio of Melan-A and SK-MEL-2 were detected by cell cloning. The changes of cell cycle of melanoma Melan-A cells were determined by flow cytometry. The levels of the Bax and Caspase3 of Melan-A apoptosis protein in melanoma cells were detected by Western blot.Results:NVP-LDE225, an inhibitor of Smo, decreased the mRNA expression of Gli1 in the melanoma cells in a dose-dependent manner, inhibited the proliferation ratio of the melanoma cells, induced apoptosis, and arrested melanoma cells in G 0 / G 1 phase. Gamma ray irradiation after NVP-LDE225 treatment further inhibited the SHH signal pathway, arrested the melanoma cells in G 2 / M phase, and further increased the inhibitory effect on melanoma cell proliferation. Conclusions:NVP-LDE225, an inhibitor of Smo, can inhibit SHH signal pathway in melanocytes, suppress cell proliferation, and further increases the effect of inhibiting cell proliferation after combining with irradiation. It can be used as a potential drug in combination with radiotherapy in the treatment of melanoma, which is worthy of further study.
RESUMO
AIM: To study the effects and mechanism of Lycium barbarum polysaccharide on the proliferation and apoptosis of osteosarcoma HOS cells. METHODS: Osteosarcoma HOS cells were divided into four groups: control group, LBP-I group, LBP-II group and LBP-III group. MTT was used to detect cell proliferation; PI single staining was used to detect cell cycle; Annexin V-FITC/PI double staining was used to detect apoptosis; Western blot was used to detect the expression of Cleaved Caspase-3, Cleaved PARP, cyclin-dependent kinase 4 (CDK4), cyclin D1, Shh and Gli1. Shh signal activator and Lycium barbarum polysaccharide treated osteosarcoma HOS cells together; the changes of cell proliferation, cell cycle and apoptosis were observed.RESULTS: Compared with the control group, the proliferation ability of LBP-I group, LBP-II group and LBP-III group decreased, the proportion of cells in G0/G1 phase increased[(51.2±4.1)% vs. (59.1±3.2)%, (66.8±2.0)%, (72.3±3.2)%, F=72.76, P<0.001], the level of apoptosis increased[(3.9±0.3)% vs. (13.2±1.2)%, (17.6±1.3)%, (24.8±2.1)%, F=364.50, P<0.001], the expression levels of Cleaved Caspase-3, Cleaved PARP protein increased, the expression levels of CDK4, cyclin D1, Shh and Gli1 decreased (P<0.05). Compared with cells not treated with Shh signal activator, the cells treated with Shh signal activator could reverse the effect of Lycium barbarum polysaccharide on the proliferation, cycle arrest and apoptosis of osteosarcoma HOS cells. CONCLUSION: Lycium barbarum polysaccharides blocked the cell cycle of osteosarcoma HOS cells, inhibited cell proliferation and promoted cell apoptosis by inhibiting Shh signaling pathway.