Application of cucurbitacine B to preparation of cancerous inhibitor of protein phosphatase 2A

A protein phosphatase and inhibitor technology, applied in the field of medicine, can solve the problems of insensitivity of anti-growth signals, tumor cell escape and apoptosis, etc., and achieve the effect of significant curative effect, clear target and broad anti-cancer application prospects.

Inactive Publication Date: 2016-11-09
HUBEI UNIVERSITY OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutations in these genes often lead to tumor cell escape from apoptosis, insensitivity to normal anti-growth signals, acquisition of self-sufficient growth signals and unlimited replication potential, as well as the ability of tissue invasion and metastasis, resulting in recurrence and drug resistance

Method used

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  • Application of cucurbitacine B to preparation of cancerous inhibitor of protein phosphatase 2A
  • Application of cucurbitacine B to preparation of cancerous inhibitor of protein phosphatase 2A
  • Application of cucurbitacine B to preparation of cancerous inhibitor of protein phosphatase 2A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Cucurbitacin B inhibits the growth of gastric cancer, breast cancer, lung cancer and glioma cells

[0017] The various types of cancer cell lines SGC7901 (gastric cancer), MCF-7 / Adr (breast cancer), NCI-H1975 (lung cancer), and DBTRG (glioma) cells in the logarithmic growth phase and human normal bronchial Epithelial cells 16-HBE, human normal gastric mucosal epithelial cells GES-1 were seeded in 96-well plates (approximately 5000 cells per well, 90μl medium), and cultured (37℃, 5% CO 2 Incubator) 24 hours later, 5 kinds of cells were treated with Cucurbitacin B (CuB) with different concentration gradient (10-1200nmol / L). After culturing for 20 and 44 hours respectively, add 10μl of 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) solution at a concentration of 5mg / ml to each well, continue Cultivate for 4 hours. After terminating the reaction, aspirate the medium, add 150 μl dimethyl sulfoxide to each well, shake at low speed to fully dissolve, ...

Embodiment 2

[0018] Example 2 Cucurbitacin B inhibits the expression of CIP2A protein in gastric cancer, breast cancer, lung cancer and glioma cells.

[0019] The specific implementation method is as follows:

[0020] Inoculate various types of cancer cell lines SGC7901 (gastric cancer), MCF-7 / Adr (breast cancer), NCI-H1975 (lung cancer) and DBTRG (glioma) cells in a 6-well plate at a certain density. Hours later, when the cell confluence reached 80%, SGC7901 cells (0nmol / L, 25nmol / L, 50nmol / L, 100nmol / L) were treated with different concentrations of CuB, MCF-7 / Adr (0nmol / L, 100nmol / L) L, 200nmol / L, 400nmol / L), NCI-H1975 cells (0nmol / L, 100nmol / L, 200nmol / L, 300nmol / L), DBTRG cells (0nmol / L, 100nmol / L, 200nmol / L, 400nmol / L) After 24 hours, the cells were lysed with 1×SDS Loading Buffer to extract total protein, and Western Blot immunoblotting experiment was performed with the same amount of protein, using anti-CIP2A and anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase) antibodies respecti...

Embodiment 3

[0021] Example 3 Cucurbitacin B inhibits the phosphatase activity of CIP2A substrate protein PP2A in gastric cancer, breast cancer, lung cancer, and glioma cells

[0022] The specific implementation method is as follows:

[0023] Inoculate various types of cancer cell lines SGC7901 (gastric cancer), MCF-7 / Adr (breast cancer), NCI-H1975 (lung cancer) and DBTRG (glioma) cells at a certain density in a 10 cm culture dish, 24 After hours, when the cell confluence reached about 70%-80%, SGC7901 cells (0nmol / L, 25nmol / L, 50nmol / L, 100nmol / L) were treated with different concentrations of CuB, MCF-7 / Adr (0nmol / L) , 100nmol / L, 200nmol / L, 400nmol / L), NCI-H1975 cells (0nmol / L, 100nmol / L, 200nmol / L, 300nmol / L), DBTRG cells (0nmol / L, 100nmol / L, 200nmol / L) , 400nmol / L), 24 hours later, use RIPA lysis solution to lyse four kinds of cells treated with different concentrations of CuB, and store the protein solution at -20°C. Take 100μg of each protein and incubate with 4μg of PP2A antibody respec...

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Abstract

The invention belongs to the technical field of medicine, and particularly relates to an application of tetracyclic-triterpene-like compound-like cucurbitacine B separated and extracted from cucurbitaceous plants as a cancerous inhibitor of protein phosphatase 2A. Cucurbitacine B is taken as a main component in the cancerous inhibitor of protein phosphatase 2A, so that the expression of the cancerous inhibitor of protein phosphatase 2A can be inhibited remarkably. The cucurbitacine B can be applied to independent administration or combined treatment, including combined applications with radiotherapy, chemotherapy, surgical treatment, other growth factor inhibitors and sexual hormone inhibitors, any tumor treatment medicament and method.

Description

Technical field [0001] The invention belongs to the technical field of medicine, and specifically relates to an application of cucurbitacin B in the preparation of a protein phosphatase 2A cancer inhibitory factor inhibitor. Background technique [0002] Tumors, especially malignant tumors, are a major disease threatening human health, and tumor treatment is a major medical problem facing humans today. Tumors are often caused by functional mutations of oncogenes and loss of functional mutations of tumor suppressor genes. Mutations in these genes often cause tumor cells to escape apoptosis, become insensitive to normal anti-growth signals, obtain self-sufficient growth signals, the potential for unlimited replication, and the ability to infiltrate and metastasize tissues, which in turn leads to recurrence and drug resistance. Therefore, tumors are actually diseases in which cell signal transduction is abnormal. Finding compounds that specifically bind to and inhibit the activity...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/575A61P35/00
CPCA61K31/575
Inventor 刘莹郭阳张亮刘雪文段超黄秋月
Owner HUBEI UNIVERSITY OF MEDICINE
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