Unlock instant, AI-driven research and patent intelligence for your innovation.

Action target spot of glucocorticoid and application thereof

A glucocorticoid and cell technology, applied in the fields of biology and medical testing, can solve problems such as unclear targets

Inactive Publication Date: 2010-06-09
谢彦晖
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the current specific targets of glucocorticoids are unclear, there are few in vitro methods to detect the strength and sensitivity of glucocorticoids

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Action target spot of glucocorticoid and application thereof
  • Action target spot of glucocorticoid and application thereof
  • Action target spot of glucocorticoid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0147] FOX O3a is a target of glucocorticoid action

[0148] Materials and methods

[0149] 6-8 weeks old Balb / C mice were purchased from the SPF animal room of Shanghai Institute of Biochemistry and Cells, Chinese Academy of Sciences

[0150] The siRNA of FoxO3a (5'GGAUGACGUCCAGGAUGAUTT 3' (forward translation strand SEQ ID NO: 2) and 5'AUCAUCCUGGACGUCAUCCTT 3' (reverse translation strand SEQ ID NO: 3) was purchased from GenePharma

[0151] Dexamethasone (DXM), purchased from sigma-aldrich company

[0152] method 1

[0153] Experimental group (group 3): Lymphocytes were obtained from the spleen of 6-8 week Balb / C mice, cultured in RPMI1640 medium and 10% fetal bovine serum, in a 6-well plate of lymphocyte suspension, about 10 per well 6 Dexamethasone was added to the cells at a concentration of 4 μM, 1 μM, and 0.1 μM;

[0154] Control group: ethanol solution without adding dexamethasone;

[0155] The cells were harvested after 6 hours, and the expression changes of FoxO3...

Embodiment 2

[0166] Sensitivity of cells to glucocorticoids in vitro

[0167] Materials and methods

[0168] Myeloma cell line SP2 / O was purchased from the cell bank of Shanghai Institute of Biochemistry and Cell, Chinese Academy of Sciences

[0169] Chronic lymphocytic leukemia cell line L1210 was purchased from the cell bank of Shanghai Institute of Biochemistry and Cell, Chinese Academy of Sciences

[0170] Gastric cancer cell line MFC was purchased from the cell bank of Shanghai Institute of Biochemistry and Cell, Chinese Academy of Sciences

[0171] Breast cancer cell line 4T1 was purchased from the cell bank of Shanghai Institute of Biochemistry and Cell, Chinese Academy of Sciences

[0172] Test group (4 groups):

[0173] SP2 / O group: culture SP2 / O in RPMI 1640 medium and 10% fetal bovine serum,

[0174] MFC group: MFC was cultured in RPMI1640 medium and 10% fetal bovine serum,

[0175] 4T1 group: 4T1 was cultured in RPMI1640 medium and 10% fetal bovine serum,

[0176] L1210 g...

Embodiment 3

[0185] Screening for glucocorticoids

[0186] Select the more sensitive tumor cell line SP2 / O, and culture SP2 / O in RPMI1640 medium and 10% fetal bovine serum,

[0187] Add the hormone to be screened, dexamethasone, prednisone, methylprednisolone, and hydrocortisone at 4 μM each to the selected tumor cell culture dish.

[0188] After 8-24 hours, the cells were collected for western blot detection, and the phospho-FoxO3a / FoxO3a ratio was selected to be low, preferably 0.2-0.3. The lower the value, the more sensitive the cells were to the hormone, and the stronger the anti-inflammatory activity of the hormone.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an action target spot of glucocorticoid and application thereof, particularly discloses an action target spot of FoxO3a which is glucocorticoid, and provides a method for screening a glucocorticoid compound, a method for screening a compound for inhibiting FoxO3a phosphorylation, a method for improving the sensitivity of a cell on glucocorticoid, a method for detecting thesensitivity of the cell on the glucocorticoid and a kit.

Description

technical field [0001] The invention relates to the fields of biology and medical testing, and relates to FoxO3a, an action target of glucocorticoid, and its application in screening drugs and detecting drug resistance of glucocorticoid. Background technique [0002] Glucocorticoids have anti-inflammatory, anti-allergic, anti-shock and anti-tumor effects, and are widely used clinically, such as in the treatment of autoimmune diseases, chronic inflammation, and hematopoietic system tumors. But there are also some diseases that are not sensitive to the application of glucocorticoids. Glucocorticoids can induce apoptosis of lymphocytes, lymphoma cells, and myeloma cells. Therefore, in resisting inflammation, inflammation can be subsided by inducing apoptosis of lymphocytes. But at the same time, glucocorticoids can protect breast cells, gastric cancer cells and other cells from hormone-induced apoptosis. Recently, it was discovered that a transcription factor protein closely r...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/02G01N33/53
Inventor 谢彦晖
Owner 谢彦晖