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Application of GSK126 to preparation of anti-osteosarcoma medicine

A technology for osteosarcoma and osteosarcoma cells, which is applied in antineoplastic drugs, drug combinations, and pharmaceutical formulations, etc.

Pending Publication Date: 2022-01-07
广州市红十字会医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that EZH2 indirect inhibitors play an inhibitory role in a variety of tumors, but there is no relevant report on the role of EZH2 direct inhibitor GSK126 in osteosarcoma

Method used

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  • Application of GSK126 to preparation of anti-osteosarcoma medicine
  • Application of GSK126 to preparation of anti-osteosarcoma medicine
  • Application of GSK126 to preparation of anti-osteosarcoma medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Cell Proliferation Experiment

[0022] (1) Experimental materials: human osteosarcoma Saos-2 cell line and human osteosarcoma U2OS cell line were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences; high-glucose DMEM was purchased from Gibco, USA; fetal bovine serum was purchased from Gibco, USA; MTS cell proliferation kit Purchased from Promega (Promega, USA); GSK126 was purchased from Selleck (China, Shanghai Bluewood Chemical Co., Ltd.); 96-well plates were purchased from Thermo (Thermo Fisher, USA).

[0023] (2) Experimental method: Human osteosarcoma Saos2 cells and human osteosarcoma U2OS cells were inoculated in 96-well plates, with about 2000 cells per well, and 100 μl culture medium, and GSK126 drugs with GSK126 concentrations of 5 μM, 10 μM, and 15 μM were added respectively. °C, 5% CO 2 After culturing for 48 hours, add 20 μl MTS respectively, continue to incubate at 37° C. for 2 hours, and detect the absorbance value at OD 490...

Embodiment 2

[0025] Embodiment 2 Immunofluorescence experiment

[0026] (1) Experimental materials: human osteosarcoma Saos-2 cell line and human osteosarcoma U2OS cell line were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences; GSK126 was purchased from Selleck (China, Shanghai Lanmu Chemical Co., Ltd.); rabbit anti-human LC3 polyclonal antibody was purchased from From CST (Cell signaling technology company in the United States); FITC fluorescent secondary antibody and DAPI staining solution were purchased from Sigma (Sigma Company in the United States); goat serum and paraformaldehyde solution were purchased from Wuhan Boster Company.

[0027] (2) Experimental method: Human osteosarcoma Saos2 cells and human osteosarcoma U2OS cells were treated with GSK126 at a concentration of 5 μM, 10 μM, and 15 μM for 48 hours, fixed with 4% paraformaldehyde solution for 15 minutes, washed 3 times with PBS, and washed with Block with 10% goat serum, then add 1:100 diluted rabbi...

Embodiment 3

[0029] Example 3 Western blot experiment

[0030] (1) Experimental materials: human osteosarcoma Saos-2 cell line and human osteosarcoma U2OS cell line were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences; GSK126 was purchased from Selleck (China, Shanghai Lanmu Chemical Co., Ltd.); rabbit anti-human LC3 polyclonal antibody, Rabbit anti-human p62 polyclonal antibody was purchased from CST (Cell signaling technology company, USA); BCA kit.

[0031] (2) Experimental method: Human osteosarcoma Saos2 cells and human osteosarcoma U2OS cells were inoculated in 96-well plates, with about 2000 cells per well, and 100 μl culture medium, and GSK126 drugs with GSK126 concentrations of 5 μM, 10 μM, and 15 μM were added respectively. °C, 5% CO 2 After culturing for 48 hours, collect human osteosarcoma Saos2 cells and human osteosarcoma U2OS cells, add an appropriate amount of cell lysis solution RIPA (containing 1 mmol / L PMSF) to each tube of cells, fully cover th...

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Abstract

The invention discloses an application of GSK126 to preparation of an anti-osteosarcoma medicine, and relates to the technical field of a new application of medicines. It is found that GSK126 can significantly inhibit osteosarcoma cell proliferation, increase expression of autophagosome and induce autophagy of osteosarcoma cells to achieve the effect of inhibiting osteosarcoma growth. The GSK126 can be used for developing a novel medicine for treating osteosarcoma, and has wide medical prospects and economic values.

Description

technical field [0001] The invention relates to the technical field of new uses of medicines, in particular to the application of GSK126 in the preparation of anti-osteosarcoma medicines. Background technique [0002] Osteosarcoma is a primary malignant tumor of mesenchymal tissue. It occurs frequently in the metaphysis of long bones in adolescents, especially the distal femur and proximal tibia. It has the characteristics of early lung metastasis, high disability rate, and high recurrence rate. Surgery and drug chemotherapy are the standard treatment options for this disease. New advances in the treatment of osteosarcoma have significantly improved the prognosis of patients, but the 5-year overall survival rate of patients with osteosarcoma metastasis or recurrence is still less than 30%. Osteosarcoma is a kind of differentiation defect disease, which is related to osteogenic differentiation-related genes and (or) epigenetic changes, and is characterized by the blockage of ...

Claims

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Application Information

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IPC IPC(8): A61K31/496A61P35/00
CPCA61K31/496A61P35/00
Inventor 熊喜峰王鹏珍汤楠张金丽赖旭东刘志河李爱国
Owner 广州市红十字会医院
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