Application of breast cancer multidrug resistant protein molecule IBP

A multi-drug resistance, breast cancer technology, applied in the field of biochemistry, can solve problems such as unsatisfactory effects, and achieve the effects of reducing expression, improving sensitivity, and reducing drug efflux capacity

Inactive Publication Date: 2017-02-08
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the results of research on the reversal of breast cancer drug resistance targeting these mechanisms are not satisfactory in clinical practice.

Method used

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  • Application of breast cancer multidrug resistant protein molecule IBP
  • Application of breast cancer multidrug resistant protein molecule IBP
  • Application of breast cancer multidrug resistant protein molecule IBP

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, induction of breast cancer drug-resistant cell line MCF-7 / ADR and cultivation of breast cancer cell line

[0025] MCF-7, MDR-MB-231, and SK-BR-3 breast cancer cell lines were purchased from ATCC cell bank in the United States and frozen in our laboratory. MCF-7 cells were cultured in 1640 medium containing 10% (v / v) fetal calf serum, and multidrug-resistant MCF-7 / ADR cells were cultured by MCF-7 parental cells continuously exposed to gradient concentrations of doxorubicin Induced in the base, wherein the concentration of doxorubicin was gradually increased from 5ng / ml to 1000ng / ml. For long-term culture of MCF-7 / ADR, 1640 medium containing 1000ng / ml doxorubicin and 10% fetal bovine serum was used at 37°C with 5% (v / v) CO 2 nourish. MDR-MB-231 and SK-BR-3 cells were cultured in DMEM medium containing 10% (v / v) fetal bovine serum.

[0026] The constructed multidrug-resistant MCF-7 / ADR cells were treated with different concentrations of doxorubicin, and th...

Embodiment 2

[0033] Example 2, Construction of MCF-7 / ADR, MDA-MB-231, SK-BR-3 stable cell lines of lentivirus-mediated IBPshRNA interference

[0034] Through bioinformatics analysis, for IRF4 binding protein (IBP) gene (Genbank: NM_022047), its amino acid sequence is shown in SEQ ID NO.11, then design RNA interference sequence according to 2 targets of IBP gene, target sequence and The interference sequences are shown in Table 2. Anneal the synthesized oligo into a double-stranded oligo sequence, and connect it into the linearized RNA interference vector LV10 to obtain the LV10 vector containing the target sequence. Correct transformants were verified by sequencing, and high-purity plasmid extraction was carried out. Then the constructed LV10 vector containing the target sequence was co-transfected into 293T cells with shuttle plasmids and packaging plasmids (pGag / Pol, pRev, pVSV-G), the cell culture supernatant was collected, the virus was concentrated by ultracentrifugation, and stored ...

Embodiment 3

[0043] Embodiment 3, construct the MCF-7 stable transfection cell line of IBP overexpression

[0044] Total RNA was extracted from human lymphocytes and reverse transcribed into cDNA. Using human lymphocyte cDNA as a template, PCR technology was used to amplify the full-length cDNA fragment of human IBP. The full-length cDNA fragment of human IBP was cloned into the eukaryotic expression vector pEGFP-C1. After sequencing and identification, the pEGFP fusion expression plasmid pEGFP-C1-IBP was successfully constructed. 1 μg of pEGFP-C1 and pEGFP-C1-IBP plasmids were respectively transfected into MCF-7 cells in a 24-well plate to obtain IBP-overexpressing MCF-7 cells. Lipofectamine2000 kit was used for transfection; the cells were digested 24 hours after transfection, and then the cell suspension was diluted at a volume ratio of 1:24 and then inoculated in a new 24-well plate, and the cell culture conditions were as described above MCF-7 cell culture conditions; add 1000 μg / ml...

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Abstract

The invention discloses an application of a breast cancer multidrug resistant protein molecule IBP, wherein the IBP has an amino acid sequence as shown in SEQ ID NO.11; studies find out that expression of the IBP in breast cancer multidrug resistant cells MCF-7/ADR is significantly higher than expression in MCF-7 parental cells, and moreover, after the expression of the IBP in the breast cancer cells is decreased, the sensibility on a variety of chemotherapy drugs is significantly enhanced; after the expression of the IBP in the MCF-7/ADR is interfered, the breast cancer drug-resistant cells can be inhibited, besides, expression and drug transport functions of MDR1 and other multidrug resistant related genes in the MCF-7/ADR are inhibited, and the drug sensitivity is increased; therefore, the IBP can be used as a multidrug resistant target gene for detection of breast cancer, and also can be used for preparing breast cancer multidrug resistant preparations or drugs.

Description

technical field [0001] The invention belongs to the field of biochemistry, and in particular relates to the application of breast cancer multidrug resistance protein molecule IBP. Background technique [0002] Breast cancer is a major disease that seriously threatens women's health and is the second leading cause of cancer-related deaths in women. Studies have found that, despite adjuvant chemotherapy, there are still a considerable number of patients with breast cancer metastasis and recurrence. According to statistics, the 5-year survival rate of metastatic breast cancer is only 26.7%. 90% of patients with metastatic breast cancer fail treatment due to chemotherapeutic drug resistance. Progressive antineoplastic drug resistance during treatment is an important issue for patients with metastatic breast cancer. More importantly, although there are a variety of anti-tumor drugs in clinic, once a drug resistance occurs during the course of treatment, most of the therapeutic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68A61K45/00A61P35/00
CPCA61K45/00C12Q2600/158G01N33/68G01N2800/7028
Inventor 杨明珍袁方胡川闽陈安李淑慧
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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