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Application of micromolecular nucleic acid miR-302 for treating or preventing testicular cancer

A technology of molecular nucleic acid and testicular cancer, applied in the field of treatment or prevention of testicular cancer

Inactive Publication Date: 2012-11-14
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are no studies on the combination of miRNA and chemotherapeutic drugs in the treatment of testicular cancer

Method used

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  • Application of micromolecular nucleic acid miR-302 for treating or preventing testicular cancer
  • Application of micromolecular nucleic acid miR-302 for treating or preventing testicular cancer
  • Application of micromolecular nucleic acid miR-302 for treating or preventing testicular cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1: experimental material preparation

[0072] 1. Cell lines, cell culture and transfection

[0073] Human malignant multiple teratoma cell NTera-2 (NT2), human prostate cancer cell DU145, breast cancer cell MDA-MB-231, liver cancer cell HepG2, and cervical cancer cell HeLa were purchased from ATCC. Each of the above cells was cultured in Dulbecco's modified Eagle's medium (DMEM) medium containing 10% fetal bovine serum (FBS, Life Technologies, Inc., Grand Island, NY, USA), 1% antibiotics (100 U / ml penicillin and 100 μg / ml streptomycin; Life Technologies); cultured at 37°C with 5% CO 2 in the incubator. NT2 cells were scraped off by cell scraping, and the remaining cells were digested with trypsin (Trypsin-EDTA solution).

[0074] LipofectamineRNAiMAX (Invitrogen) transfection reagent was used to transfect the above cells with small RNA. Unless otherwise specified, small interfering RNAs are 50nM siRNAs, microRNA mimics and their controls are 100nM, and m...

Embodiment 2

[0086] Example 2: Analysis of the functions of miR-302a and its related genes by means of cell biology experiments

[0087] 1. Western blotting

[0088] Reagent preparation:

[0089] 1) RIPA lysis buffer: 50mMTris-HCl (pH 7.4), 150mMNaCl, 1% TritonX-100, 1% sodium dodecyl sulfate, 1% sodium deoxycholate, 1mM EDTA,

[0090] and add as needed:

[0091] Protease inhibitor: EDTA-free protease inhibitor cocktail (Roche) and 1mM benzene

[0092] Methanesulfonyl fluoride (PMSF)

[0093] Phosphatase inhibitor: 5mM sodium orthovanadate

[0094] 2) Protein electrophoresis separation gel and stacking gel

[0095]

[0096] 3) Reduced 5×SDS loading buffer: Tris HCl (pH 6.8) 250mM, β-mercaptoethanol 5%, SDS 10%, bromophenol blue 0.5%, glycerol 50%

[0097] 4) Tris-glycine electrophoresis buffer: 25mM Tris, 250mM glycine (pH 8.3), 0.1% SDS;

[0098] 5) Transfer buffer: Tris 3.03g / L, Glycine 18.8g / L, SDS 1g;

[0099] 6) 10× Ponceau dye solution: Ponceau S 2g, trichloroacetic acid 3...

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PUM

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Abstract

The invention relates to endogenic non-coding microRNA (micro Ribonucleic Acid), a medicine composition containing the microRNA, and new application of the medicine composition for treating or preventing the testicular cancer. According to the invention, the microRNA belonging to the family member (miR-302) of the microRNA-302 and platinum-type anti-tumour medicines are utilized jointly, so that the treatment effect of the platinum-type anti-tumour medicines for the testicular cancer can be obviously improved, and simultaneously, the administration dose, the administration time course and the side effects of the platimun-type anti-tumour medicines can be reduced. The invention also improves a molecular mechanism of the platinum-type anti-tumour medicines for treating the testicular cancer and develops a new treatment target spot for treating the testicular cancer, thereby having a significant value.

Description

technical field [0001] The present invention relates to an endogenous non-coding small RNA, a pharmaceutical composition containing it, and its new application for treating or preventing testicular cancer. Specifically, the present invention relates to a novel use of microRNA-302 family members (miR-302) for treating or preventing testicular cancer. Background technique [0002] Testicular cancer is a relatively rare type of cancer in the general population but one of the most common malignancies in young men aged 15-35 years (1). In recent decades, the global incidence of testicular cancer has gradually increased, especially in some western countries, the incidence is increasing at a rate of 1% to 2% per year (2-3). The incidence of testicular tumors varies around the world. Blacks seldom have this disease. The incidence is about 1 / 100,000 men in Eastern Europe and Asia, and 2 / 10 to 6.3 / 100,000 men in Europe and America. In my country, the morbidity and mortality are abou...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P35/00A61K33/24A61K31/282
Inventor 孙斐刘琳
Owner UNIV OF SCI & TECH OF CHINA
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