Application of miR-1914* and miR-1915

A technology of mir-1915 and mir-1914, applied in the field of microRNA, can solve the problems in the preliminary stage and achieve the effect of inhibiting the metastasis of colorectal cancer

Inactive Publication Date: 2015-03-04
FUDAN UNIV SHANGHAI CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the research on the biological role of microRNA in the process of tumorigenesis and development has made great progress, but there are still many problems to be solved, and many studies are still in the preliminary stage.

Method used

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  • Application of miR-1914* and miR-1915
  • Application of miR-1914* and miR-1915
  • Application of miR-1914* and miR-1915

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 (Differential expression of microRNA in human colorectal cancer chemotherapy-resistant and sensitive plasma)

[0032] 1. Experimental preparation.

[0033] 1. Research object.

[0034] Patients with stage IV colorectal cancer undergoing resection of the primary lesion in the Department of Colorectal Surgery, Cancer Hospital Affiliated to Fudan University were selected. The patients did not receive radiotherapy and chemotherapy before the operation, but received standardized first-line XELOX chemotherapy after the operation. Diagnosis, grading, and staging of colorectal cancer organizations were carried out in accordance with the National Colorectal Cancer Cooperative Group colorectal cancer diagnosis and treatment guidelines and the AJCC / UICC TNM staging standards. A total of 100 subjects were studied, which were divided into 2 groups: stage IV colorectal cancer drug-resistant group (n=57), and stage IV colorectal cancer effective group (n=43). The deta...

Embodiment 2

[0075] Example 2 (in vitro experiments and target gene functions of miR-1914* and miR-1915)

[0076] 1. Establishment and selection of drug-resistant cell lines.

[0077] 1. Using the existing drug-resistant cell line culture method, the LoVo, SW620, and HCT-116 cell lines provided by the Chinese Academy of Sciences were used for culture, and the drug 5-FU and Oxaliplatin (Oxaliplatin) were used to treat each cell line. For induction, the initial concentrations of 5-Fu and Oxaliplatin were both 0.01ug / ml, and the drug concentration was increased by 1 times every time the cells were passaged three times, respectively:

[0078] 0.01 / 0.02 / 0.04 / 0.08 / 0.16 / 0.32 / 0.64 / 1.28 / 2.56 / 5.12 / 10.24 / 20.48 / 40.96.

[0079] 2. Use CCK-8 to detect cell IC50.

[0080] 3. Analyze and compare the IC50 changes of drug-resistant strains and their parental cell lines to two chemotherapeutic drugs.

[0081] 4. Use real time PCR to detect the miRNA content in drug-resistant cell lines.

[0082] The te...

Embodiment 3

[0183] Example 3 (in vivo experiments of miR-1914* and miR-1915)

[0184] 1. Experimental preparation.

[0185] 1. Cell lines.

[0186] Self-induced HCT116 / 5-Fu / Oxa (same as Example 2).

[0187] 2. Animals.

[0188] 30 nude mice, pure BALB / C (nu / mu) strain, weighing 18-20 g, all male, 4-6 weeks old, provided by the Animal Laboratory Center of Fudan University School of Medicine, raised strictly under SPF conditions, free Intake of water and food, each experiment was carried out in a sterile ultra-clean workbench.

[0189] Second, the experimental steps.

[0190] 1. Establishment and observation of xenograft nude mouse model.

[0191] HCT116 / 5-FU / OXA and its transgenic cell lines in the logarithmic growth phase were taken, and the cell suspension was prepared by trypsinization, and the cells were resuspended with fetal bovine serum, and the cell concentration was adjusted to 107 cells / mL. Each nude mouse was placed in the left 0.1 mL was injected subcutaneously under th...

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Abstract

The invention relates to application of miR-1914* and/or miR-1915 as drug resistance markers of a colorectal cancer chemotherapy drug as well as application of the miR-1914* and/or the miR-1915 in preparing a drug with the functions of inhibiting transfer, invasion, apoptosis and proliferation of the colorectal cancer. According to the invention, the miR-1914* and the miR-1915 exist in blood plasma, can be detected, are in a down-regulated trend in the blood plasma of a colorectal cancer drug resistant patient, have relative tissue specificities, are related to colorectal cancer first-line chemoresistance, and meanwhile also influence patient prognosis. The miR-1914* and the miR-1915 can be used singly or used together, and play a relatively important role in the colorectal cancer first-line chemoresistance, and particularly, the effect of combined use is better.

Description

technical field [0001] The design of the invention relates to the use of a microRNA, which belongs to the field of biotechnology. Background technique [0002] Colorectal cancer (CRC) is the general term for colon cancer and rectal cancer. Colorectal cancer refers to the malignant lesion of the colorectal epithelium under the action of various carcinogenic factors such as environment or genetics. It has a poor prognosis and a high mortality rate. The incidence of colorectal cancer ranks third among male malignant tumors worldwide. According to the data provided by the International Organization for Research on Cancer (IARc), in Asia, the death rate in East Asia is 120.1 / 100,000. With the improvement of living standards and changes in eating habits in my country, the overall incidence is also on the rise. In recent years, the incidence of colorectal cancer in Shanghai has increased by 4.2% every year, from the seventh place in the malignant tumors to the third place. Studi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C07K14/47A61K48/00A61P35/00
CPCA61K31/713C12N15/113C12N2310/141C12Q1/6886C12Q2600/106C12Q2600/158C12Q2600/178
Inventor 胡劲松蔡三军
Owner FUDAN UNIV SHANGHAI CANCER CENT
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