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Application of synuclein gamma (SNCG) gene in diagnosis and hormone therapy of breast cancer

A technology of hormone therapy and anti-estrogen therapy, which is applied in the field of diagnosis and hormone therapy to achieve accurate diagnosis

Inactive Publication Date: 2012-07-18
XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the application of SNCG gene in the diagnosis and hormone therapy of breast cancer.

Method used

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  • Application of synuclein gamma (SNCG) gene in diagnosis and hormone therapy of breast cancer
  • Application of synuclein gamma (SNCG) gene in diagnosis and hormone therapy of breast cancer
  • Application of synuclein gamma (SNCG) gene in diagnosis and hormone therapy of breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Construction and packaging of lentiviral vector

[0023] 1. Experimental materials

[0024] (1) Main equipment

[0025] YJ-875 ultra-clean workbench, C0 2 Incubator 3111, -80°C low-temperature refrigerator, LDZX-40BI pressure steam sterilizer, TGL-16G desktop high-speed centrifuge, Nanopure ultrapure water instrument, flow cytometer, AA-200 electronic balance, carton microscope CCD camera system, CK40-F200 inverted microscope, XSP-2C biological microscope, fluorescent quantitative PCR instrument, western blot electrophoresis instrument, 90-2 magnetic stirrer, 10I, tm polyester filter membrane, micro pipette gun Wait.

[0026] (2) Main drugs and reagents

[0027] Glutamine (L-Glutamine), dimethyl sulfoxide (DMSO), trypsin, fetal bovine serum, DMEM culture powder, mouse anti-human monoclonal PE-CD133 / 1, mouse anti-human ERalpha receptor antibody, small Mouse anti-human BCRP antibody, rabbit anti-human SNCG antibody, alpha-actin antibody-SC-8432, MTT, poly...

Embodiment 2

[0083] Example 2 MTT method to measure the proliferative activity of cells

[0084] 1. Experimental materials (see Example 1)

[0085] 2. Experimental steps:

[0086] MTT solution configuration: Weigh 0.5 g of MTT, dissolve in 100 ml of phosphate buffered solution (PBS), filter with a 0.22 μm filter membrane to remove bacteria in the solution, and store at 4°C in the dark.

[0087] 1) The cells were grouped according to different experiments, and after transfection with different vectors, each well was cultured with cell culture medium for 2 hours, 24 hours, 48 ​​hours, and 72 hours, and three replicate holes were selected for each group.

[0088] 2) After taking out the incubator, add 500μl cell culture medium and 50μl MTT solution (5mg / ml) to each well. Incubate for 4 hours in a 37°C incubator.

[0089] 3) Carefully suck out the cell culture medium, add 500 μl DMSO, and shake for 10 minutes to fully dissolve the crystals.

[0090] 4) Take out 100 μl from each well and ...

Embodiment 3

[0093] Example 3 Cell cycle detection by flow cytometry

[0094] 1. Experimental materials (see Example 1)

[0095] 2. Experimental steps:

[0096] 1) Take cells from each experimental group, digest with trypsin, centrifuge, and wash with PBS;

[0097] 2) The above cells were made into single-cell suspensions, and the number of cells was adjusted to 1×10 6 a / L.

[0098] 3) Add ice-cold 70% ethanol to fix, 4°C, 1-2 hours.

[0099] 4) Centrifuge to discard the fixative, and resuspend in 3ml PBS for 5min.

[0100] 5) Add 150 ul RNaseA (diluted in 250-500 ug / ml PBS) to resuspend the cells and digest at 37°C for 30 minutes.

[0101] 6) Filter once with a 400-mesh sieve, centrifuge at 500-1000r / min for 5min, discard PBS.

[0102] 7) Stain with 1ml PI (50ug / ml) staining solution, 4°C in the dark for 30min.

[0103] 8) Flow cytometry detection, PI is excited by a 488nm argon ion laser, received by a 630 band-pass filter, and passed through the FSC / SSC scatter diagram.

[010...

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PUM

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Abstract

The invention relates to the application of synuclein gamma (SNCG) gene in diagnosis and hormone therapy of breast cancer, and the hormone therapy of breast cancer can be influenced by SNCG gene through adjusting the activated state of estrogen receptor (ER). The invention also relates to the application of the SNCG gene in the preparation of products for diagnosing the breast cancer. The invention has the advantages that the SNCG gene expression can be used for adjusting ER and hormone therapy sensibility, so that a new thought is created for the hormone therapy of the breast cancer; the SNCG gene expression can be used for adjusting ER and hormone therapy sensibility, so that the possibility of developing a new medicine for treating the breast cancer can be improved; and the SNCG gene can be used as a specific marker gene for diagnosing the breast cancer, so that the breast cancer can be more accurately and rapidly diagnosed.

Description

technical field [0001] The invention relates to an application of SNCG gene, in particular to the application of SNCG gene in the diagnosis and hormone therapy of breast cancer. Background technique [0002] Breast cancer is the most common and important breast disease, and its incidence rate is increasing year by year. Take Shanghai as an example. In 1972, the incidence rate of breast cancer among women in Shanghai was 17 per 100,000 people. In 1992, it rose to every 100,000 people. In 2000, it rose rapidly to 56.2 per 100,000 people. That is to say, the rate of increase in the 8 years from 1992 to 2000 exceeded the rate of increase in the 20 years from 1972 to 1992, making it the malignant tumor with the highest incidence rate in women, seriously endangering the physical and mental health of women. As an estrogen-dependent tumor, the research on the status of estrogen and estrogen receptor (ER) in the pathogenesis, progression and treatment of breast cancer has been paid ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K48/00A61P35/00
Inventor 吴克瑾翁子毅陆云姝林清王永坤
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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