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Use of miR-100 inhibitor in cancer metastasis reduction

A technology of use and reagent, applied in the field of medicine

Active Publication Date: 2016-12-21
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Many cytokines in the tumor microenvironment, including TNF-α, IL-4, IL-10, and TGF-β, etc., can participate in the phenotype regulation of macrophages and affect the expression of microRNA, but the tumor microenvironment is related to tumor There are few reports on the regulation of microRNA function in macrophages

Method used

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  • Use of miR-100 inhibitor in cancer metastasis reduction
  • Use of miR-100 inhibitor in cancer metastasis reduction
  • Use of miR-100 inhibitor in cancer metastasis reduction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: Construction of mammary gland tumor mouse model

[0044] 1. Experimental animals

[0045] BALB / c female mice were used in the mammary gland tumor model mouse experiment, purchased through the Experimental Animal Center of the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences. All experimental mice were raised and vaccinated in SPF grade animal room. The temperature of the animal room was controlled at 19-24°C, the humidity was maintained at 50%-60%, and light and dark were alternated for 12 hours. The mouse feed and litter were all sterilized by high temperature, and the drinking water was sterilized by high temperature and high pressure. During the experiment, the state of the mice was observed in time, the litter was changed twice a week, and the feed and drinking water were replenished daily.

[0046] 2. Model building method

[0047] In this example, a mouse 4T1 cell line was used to establish a tumor model, which was describe...

Embodiment 2

[0072] Example 2: Transcriptome and microRNA sequencing and joint analysis of tumor-associated macrophages

[0073] After extracting RNA from tumor-associated macrophages (TAM) and spleen macrophages (SM) obtained in Example 1, RNA quality inspection was performed, and sequencing was arranged after passing the quality inspection. The selected sequencing platform was illumina Hiseq2500.

[0074] The amount of sequencing data per sample for microRNA is 20M (Table 1), and the amount of sequencing data for each sample of transcriptome mRNA is 10G. For the sequencing data obtained by microRNA and transcriptome, calculate the average quality of the base sequencing quality of each position on the read, and calculate the average quality value of each read, and count the number of reads with corresponding quality values, through the cumulative distribution The curve looks at the average mass distribution of the data to obtain a mass distribution map.

[0075] The q30% of microRNA (the...

Embodiment 3

[0091] Example 3. miR-100 is highly expressed in tumor-associated macrophages

[0092] Next, two different induction methods of tumor supernatant culture and tumor cell co-culture were carried out in vitro:

[0093] Tumor supernatant culture test group: the culture supernatant of 4T1 tumor cells was collected, mixed with fresh medium at a ratio of 1:2, and used as a conditioned medium for culturing RAW264.7 cells. Tumor cell co-culture experiment group: RAW264.7 cells were co-cultured with 4T1 cells, using a transwell chamber. Control group: only mouse monocyte RAW264.7.

[0094] Two methods induced mouse monocyte RAW264.7 cells to have the phenotype of tumor-associated macrophages. RT-PCR detected that the expression of Arg1 was significantly up-regulated in the two induced cells ( figure 2 ). The highly expressed microRNAs in eight candidate TAMs were verified again by qPCR in two induced cells ( image 3 , Figure 4 ). According to the verification results, most of ...

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Abstract

The invention relates to a use of an miR-100 inhibitor in cancer metastasis reduction, and concretely relates to a use of a reagent for determining the miR-100 expression level in the preparation of breast cancer diagnosis reagents, and a use of the miR-100 inhibitor, and a use of the miR-100 inhibitor or an miR-100 antagonist in the preparation of medicines for treating breast cancer. Up-regulation of miR-100 in in-vivo tumor related macrophages and in-vitro induced M2 type macrophages is induced by tumor micro-environment. In in-vivo test, the microRNA antagonist knocks down the expression of the miR-100 in mouse mammary gland tumor tissues, and a case that in-situ tumor growth has no statistic difference, the quantity of lung metastasis tubercles is obviously down-regulated and the mammary gland tumor metastasis is inhibited in an experiment group of the miR-100 down-regulation in the tumor related macrophages is found.

Description

technical field [0001] This application relates to the field of medicine. Specifically, it relates to the use of microRNA in the diagnosis and treatment of breast cancer. Background technique [0002] People are increasingly aware that the study of tumors should not only stay on tumor cells in isolation, but should consider the tumor and its environment as a whole, that is, the tumor microenvironment (Tumor Microenvironment, TME). The cells in the TME mainly include tumor cells, fibroblasts, adipocytes, and infiltrating inflammatory cells, among which the inflammatory cells mainly include infiltrating immune lymphocytes, macrophages, and granulocytes. The interaction between tumor cells and immune cells in the TME is an important factor promoting tumor growth and invasion (Gajewski, T.F., H. Schreiber and Y.X. Fu, Innate and adaptive immune cells in the tumor microenvironment. Nature Immunology, 2013.14 (10 ): p.1014-1022). [0003] Tumor-associated macrophages (Tumor Ass...

Claims

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Application Information

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IPC IPC(8): C12Q1/68A61K48/00A61K31/7088A61P35/04
CPCA61K31/7088A61K48/00C12Q1/6886C12Q2600/178
Inventor 罗云萍王威
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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