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Novel application of hsa-miR-185

A molecular and product technology, applied in the new application field of hsa-miR-185, can solve the problems such as the unclear function of miR-185

Active Publication Date: 2011-06-22
INST OF MODERN PHYSICS CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The function of miR-185 is still unclear

Method used

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Experimental program
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Effect test

Embodiment 1

[0059] Embodiment 1, cell transfection

[0060] 1. Preparation and validation of transfected miR-185 analogs, inhibitors and negative controls in 786-O cells:

[0061] 1. Plating cells: Inoculate 786-O cells in the logarithmic growth phase in a 12-well plate, the number is 8×10 5 / well, transfected the next day.

[0062] 2. Transfection: 1 μL of miR-185 analog (or miR-185 inhibitor or miR-185 analog negative control or miR-185 inhibitor negative control) (concentration 30 μM) was mixed in 100 μL of serum-free opti -MEM medium, 2 μL Lipo2000 mixed with 100 μL serum-free opti-MEM medium, incubated at room temperature for 10 minutes; mixed the above premixed solution and incubated at room temperature for 15 minutes; discarded the cell culture medium in the well plate, added serum-free opti- MEM (Gibico) medium, add 800 μL per well, add 200 μL of transfection mixture, 37 ° C, 5% CO 2 Culture in a cell incubator; change 1640 culture medium (10% serum) after 4 hours for culture, ...

Embodiment 2

[0090] Example 2, miR-185 enhances the sensitivity of tumor cells to radiation

[0091] 1. After irradiation, the expression level of miR-185 is generally down-regulated in tumor tissues and cell lines

[0092] Kidney cancer tissues and paracancerous tissues of renal cancer were taken from the operating room of the Second Affiliated Hospital of Lanzhou University; gastric cancer tissues and paracancerous tissues of gastric cancer were taken from the operating room of the First People's Hospital of Lanzhou City.

[0093] 823 was purchased from China Center for Type Culture Collection; 803 was purchased from Shanghai Institute of Cells, Chinese Academy of Sciences.

[0094] (1) The radiation dose is 4Gy

[0095] The radiation detection experiment of the organization: the X-ray irradiation device is 8MV X-rays produced by the electron linear accelerator (CL2100, Varian, Germany) of the Lanzhou General Hospital Radiation Therapy Center of Lanzhou Military Region, the LET is about...

Embodiment 3

[0129] Example 3, miR-185 inhibits tumor growth (tumor formation experiment in mice)

[0130] (1) Tumor formation ability experiment of 786-O in SCID / NOD mice

[0131] Non-obese diabetes-SCID mice (NOD / SCID) mice have combined T and B lymphocyte immunodeficiency, low NK cell activity, and no circulating complement, macrophages, and antigen-presenting cells In recent years, it has become the best research model for human tumor xenografts. SCID / NOD mice were purchased from Beijing Huafukang Biotechnology Co., Ltd., 6 weeks old, weighing 18-22 g, and raised in the SPF-level animal breeding room of the Pharmacology and Toxicology Experimental Center of Gansu Cancer Hospital. The mice were randomly divided into 6 groups with 3 mice in each group. Subcutaneous axillary injection, the number of inoculated cells is 2×10 5 / 200 μL, observe and record the tumor growth every day until the tumor grows to an appropriate volume, execute, take pictures, weigh the tumor, and store the tumo...

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Abstract

The invention relates to novel application of hsa-miR-185. The novel application is application of miR-185 in preparing products with function of enhancing the sensitivity of cells and / or tissues and / or organisms to radiation, or preparing products with function of treating and / or preventing tumors, or preparing products inhibiting signal channels activated or mediated by ATR genes, products inhibiting signal channels activated or mediated by ATR proteins, products inhibiting the ATR genes and / or products for inhibiting the ATR proteins. Experiments prove that: the miR-185 can directly inhibit tumor growth and enhance the sensitivity of cells to the radiation. Further experiments prove that: the functions of the miR-185 are realized by inhibiting the expression of the ATR genes in the cells. Therefore, the miR-185 is used for preparing medicines for treating tumors and radiosensitizers, and has important significance for treating the tumors and wide application prospect.

Description

technical field [0001] The present invention relates to a new application of hsa-miR-185. Background technique [0002] Tumor is a common and frequently-occurring disease that seriously threatens human life and health. It accounts for the top three disease-related mortality in many countries and regions, and its morbidity and mortality are still increasing year by year. The current main treatment methods include surgery, radiotherapy and chemotherapy. According to the data of the World Health Organization (WHO) in 1998, the total cure rate of tumors is about 45%, and 80% of patients have received radiotherapy to varying degrees, but the radiation resistance of tumors seriously affects the therapeutic effect of radiotherapy. [0003] Radiation therapy is one of the very important tumor treatment methods, but the cure rate is still low. In addition, the current radiotherapy devices mainly include γ-rays, X-rays, and the like. The mechanism of action of these conventional ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63A61K48/00A61P35/00
Inventor 周光明苏锋涛何进鹏徐瑚珊丁楠朱佳贇胡文涛
Owner INST OF MODERN PHYSICS CHINESE ACADEMY OF SCI
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