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Method for reversing drug resistance of breast cancer by using miR-487a

A mir-487a, mir-487amimic technology, applied in the application field of miR-487a in reversing drug resistance of breast cancer cells, can solve the problem of unclear reversion of breast cancer cells

Inactive Publication Date: 2013-08-28
魏敏杰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In the present invention, we can use biological information analysis to obtain that there is a target for miR-487a to regulate BCRP expression in BCRPmRNA-3'UTR, but whether miR-487a regulates the expression of BCRP and thus reverses the resistance of breast cancer cells drug is not yet known

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  • Method for reversing drug resistance of breast cancer by using miR-487a
  • Method for reversing drug resistance of breast cancer by using miR-487a
  • Method for reversing drug resistance of breast cancer by using miR-487a

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Embodiment approach

[0043] A method for reversing drug resistance in breast cancer using miR-487a, the method comprising:

[0044] (1) Using miR-487a mimic, miR-487a mimic, inhibits BCRP expression and drug transport function in breast cancer drug-resistant cells MCF-7 / MX, and increases sensitivity to BCRP transport substrate mitoxantrone hydrochloride MX ;

[0045] (2) Using miR-487a miR-487a agmir liposome, a mimetic of miR-487a, to target and inhibit the expression of BCRP in breast cancer xenografts induced by MCF-7 / MX, and increase the drug sensitivity of somatic tumor cells to MX ;

[0046] (3) Using miR-487a inhibitor miR-487a inhibitor to up-regulate the BCRP expression and drug transport function of drug-sensitive cell MCF-7, and increase drug resistance to MX.

[0047] Below in conjunction with specific embodiment, the present invention is described in further detail, but not limitation of the present invention:

Embodiment 1

[0048] Example 1: miRNA chip analysis of miRNAs expression profile difference in breast cancer sensitive cell MCF-7 and drug-resistant cell MCF-7 / MX;

[0049] The human breast cancer cell MCF-7 used in the present invention was purchased from the ATCC cell bank in the United States, and was cryopreserved by our laboratory. MCF-7 / MX was induced by our laboratory. The medium is DMEM medium (high sugar) containing 10% fetal bovine serum, which is supplemented with 100 U / ml penicillin and 100 U / ml streptomycin, 5% CO 2 , cultured in a 37°C incubator. The difference in the expression profile of miRNAs between the sensitive breast cancer cell MCF-7 and the drug-resistant cell MCF-7 / MX was analyzed by miRNA microarray, and 48 miRNAs were found to be differentially expressed (see figure 1 ), among which miR-21, miR-302a, miR-302b, miR-487a, etc. were all lowly expressed in MCF-7 / MX.

Embodiment 2

[0050] Embodiment 2: Real-time PCR detects the expression difference of miR-487a in MCF-7 and MCF-7 / MX;

[0051] After the MCF-7 and MCF-7 / MX cells were cultured for 24 hours, the cells were collected, and the total RNA was extracted according to the steps of the RNA extraction kit (Bioteke, RP5301), and the concentration and quality of the RNA were determined by a UV spectrophotometer. Using SYBR real-time PCR kit (Takera, DRR036S), according to 10 μl system (DEPC H 2 (0.45 μl, 5*buffer 2 μl, RRI 0.25 μl, M-MLV 0.3 μl, RT-primmer 1 μl, dNTP 1 μl, RNA 5 μl), the reaction conditions are: 30°C, 10min; 42°C, 1h; 85°C, 5min; 5 ℃, 5min; 4℃, 2h, cDNA was obtained by reverse transcription. Then carry out real-time PCR experiment according to 25μl system (deionized water 9μl, 2*SYBRgreen12.5μl, Rox0.5μl, upstream primer 0.5μl, downstream primer 0.5μl, cDNA 2μl), the reaction conditions are: 95℃, 2min; 95℃ , 15s; 60°C, 30s, 40 cycles; 95°C, 1min; 55°C, 30s; 95°C, 30s. The experiment...

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Abstract

The invention belongs to the field of medicines for treating cancer, and in particular relates to a method for reversing drug resistance of breast cancer by using miR-487a. The method comprises the following steps: (1) inhibiting BCRP (Breast Cancer Resistance Protein) expression and drug transport function in drug resisting cells MCF-7 / MX of breast cancer by means of a stimulant miR-487amimic of miR-487a, and increasing sensitivity to a BCRP transfer substrate mitoxantrone hydrochloride; (2) performing targeted inhibition on BCRP expression in transplantation tumor of nude mouse with breast cancer induced by MCF-7 / MX by means of a stimulant miR-487aagmir liposome, and increasing sensitivity to MX by in vivo cancer cells; and (3) up-regulating BCRP expression and drug transport function of drug susceptible cells MCF-7 by means of an inhibitor miR-487ainhibitor of miR-487a, and increasing the drug resistance to MX. The method is used for reversing drug resistance of breast cancer by using miR-487a.

Description

Technical field: [0001] The invention provides a field of drugs for treating tumors, and particularly relates to an application of miR-487a in reversing drug resistance of breast cancer cells. Background technique: [0002] Breast cancer is one of the most common malignant tumors in women, seriously affecting women's physical and mental health and even threatening their lives. Chemotherapy is a common method for the treatment of breast cancer, but the problem of drug resistance during chemotherapy has seriously affected the clinical efficacy and prognosis of patients. Breast cancer resistant protein (BCRP), which is closely related to drug resistance in breast cancer, is a transmembrane half-transporter isolated from the drug-resistant human breast cancer cell line MCF7 / AdrVP by mRNA fingerprinting. Its gene is located on human chromosome 4q22, encoded by the ABCG2 gene, composed of 655 amino acid residues translated from 2.4kb mRNA, containing an ATP binding domain and a h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P35/00
Inventor 魏敏杰何苗马孟涛赵琳白雪峰于兆进孙明立陈秋晨
Owner 魏敏杰
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