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New use for ER (estrogen receptor) antagonist

An antagonist and tumor cell technology, applied in the field of biomedicine, can solve the problems of chemotherapy failure, poor curative effect of breast cancer patients, and deterioration of patients' condition, and achieve the effect of wide application and good reversal of tumor drug resistance

Inactive Publication Date: 2012-06-27
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the gradual application of chemotherapy regimens based on paclitaxel anticancer drugs, the cure rate of breast cancer patients has been considerably improved, but there are still a considerable number of breast cancer patients with poor curative effect, the main reason is due to tumor resistance. The drug properties caused the failure of chemotherapy, and even led to the deterioration of the patient's condition and death

Method used

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  • New use for ER (estrogen receptor) antagonist
  • New use for ER (estrogen receptor) antagonist
  • New use for ER (estrogen receptor) antagonist

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The paclitaxel-resistant breast cancer MCF-7 cell line (MCF-7 / PTX) was established by simulating the clinical administration method, and the cell drug resistance was analyzed by MTT method. MCF-7 cell line was purchased from ATCC Company.

[0027] Cell culture: The drug-resistant cell line MCF-7 / PTX was established by conventional paclitaxel intermittent administration method. The cells were divided into four groups, and paclitaxel was added to the cell culture in the logarithmic growth phase to a final concentration of 10 nM, 50 nM, 200 nM, and 1000 nM, respectively. Many cells detached and died, and the adherent surviving cells grew slowly, and were passaged when the cells reached 80% saturation. They were administered weekly with the same drug concentration, continuously stimulated 10 times, and cultured for 6 months without drug.

[0028] Light absorbance value detection: Digest and centrifuge the cells in the logarithmic growth phase, prepare a single cell suspen...

Embodiment 2

[0032] Quantitative PCR and western blot were used to detect the expression of MDR1 and ER in drug-resistant and sensitive strains.

[0033] RT-PCR Cultivate sensitive MCF-7 cells and MCF-7 / PTX cells (established in Example 1), extract total RNA from the cells, use reverse-transcribed cDNA as a template, use primer5 to design primers, and perform PCR reaction.

[0034] The total protein extracted from cells was used to detect the expression of ER protein by Western blot. Before adding the sample, use Bio-Rad’s DC Protein Quantitative Detection Kit to measure the protein concentration of the sample; the amount of the sample added during electrophoresis is 20 μg, and the Tris-glycine-SDS electrophoresis buffer is used for electrophoresis at 120V for 1.5-2 hours; after electrophoresis, use The separated protein bands were transferred to PVDF membrane (Biorad) by electrotransfer; the membrane was incubated in TBST-buffer (20mM Tris-HCl, 150mM NaCl, 0.1% Tween-20, pH 7.6) containin...

Embodiment 3

[0037] Construct ER overexpression plasmids and interference plasmids, and transfect sensitive MCF-7 cells and drug-resistant MCF-7 / PTX cells, respectively.

[0038] Using conventional molecular cloning techniques, construct ER overexpression plasmids and interference plasmids, among which, the overexpression plasmid uses pcDNA3.1 (+) vector (Invitrogen Company), the interference plasmid uses pSUPER vector (OligoEngine Company), and the constructed plasmid is used in conventional They were transferred into MCF-7 cells and MCF-7 / PTX cells under the mediation of liposomes, and western blot was used to monitor the level of ER changes, while MTT and Real-Time PCR were used to detect drug resistance-related indicators.

[0039] The constructed ER overexpression plasmid pcDNA3.1(+)-ERα was verified to be correct by sequencing, and the results of PCR identification were as follows: image 3 shown. The constructed ER interference plasmid pSUPER-ERα-shRNA was verified to be correct by...

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Abstract

The invention discloses a new use for an ER (estrogen receptor) antagonist, wherein the new use is an application in preparation for a medicine used for treating the paclitaxel resistance of tumour cells, the tumour is breast cancer, and the ER antagonist is ER antagonist ICI182,780. Experiment confirms that the high-level expression of ER (estrogen receptor) is an important related index of the medicine resistance of breast cancer cells; the ER antagonist has the advantages of inhibiting the expression level of ER protein and blocking the influence of estrogen on the occurrence and development of breast cancer; and the new use for the ER antagonist exerts good function of reversing the medicine resistance of tumour in paclitaxel-resistant breast cancer cell model, and has wide application prospect in medicine used for treating the paclitaxel resistance of human breast cancer.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a new application of an ER antagonist. Background technique [0002] Breast cancer is the most common malignant tumor in women, and millions of women around the world suffer from breast cancer every year. In my country, breast cancer has become one of the fastest-rising malignant tumors, and now ranks first in the mortality rate of female malignant tumors. Chemotherapy is the main adjuvant therapy for breast cancer, and paclitaxel compounds have been effective chemotherapy drugs for breast cancer in the past ten years. With the gradual application of chemotherapy regimens based on paclitaxel anticancer drugs, the cure rate of breast cancer patients has been considerably improved, but there are still a considerable number of breast cancer patients with poor curative effect, the main reason is due to tumor resistance. Drug properties caused the failure of chemotherapy, and ev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/5685A61P35/00A61P15/14
Inventor 孙玉洁时俊锋韩晓
Owner NANJING MEDICAL UNIV
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