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New use of microRNA 145

A tumor, coding gene technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, medical preparations containing active ingredients, etc., can solve problems such as lack of research reports

Active Publication Date: 2014-06-04
普华赛尔生物医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the existing research on microRNA 145 mainly focuses on its ability to inhibit the proliferation of cancer cells, and there are almost no research reports on other methods of microRNA 145.

Method used

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  • New use of microRNA 145
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  • New use of microRNA 145

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Acquisition of metastatic colorectal cancer cell lines stably and highly expressing microRNA 145

[0033] 1. Obtainment of colorectal cancer cell lines

[0034] Colorectal cancer cell line HCT-8 (ATCC, CCL-244), its medium is the 1640 medium (Invitrogen, 11875 -093).

[0035] A. Cell recovery

[0036] (1) Take out the cryopreservation tube of HCT-8 (ATCC, CCL-244), put it into a water bath at 37°C-40°C immediately, and shake it quickly until it melts completely.

[0037] (2) Transfer the cell suspension into 10ml cold PBS, put it into a 15ml centrifuge tube, centrifuge the cells with a horizontal centrifuge (1000 rpm, 8 minutes) and discard the supernatant.

[0038] (3) Suspend the cells with an appropriate amount of fresh medium, transfer the cell suspension into a culture bottle, and culture in a carbon dioxide incubator at 37°C.

[0039] B. Cell culture

[0040] Put the colorectal cancer cell line into the culture medium, at 37°C, 5% CO 2 According t...

Embodiment 2

[0069] Embodiment 2, HCT-8 high Functional study of cell lines stably and highly expressing microRNA 145

[0070] 1. Cell growth curve

[0071] (1) 24 hours after transfection, the HCT-8 obtained by Example 1 in a good growth state high , Digested by general subculture method, made into cell suspension. Counting, accurately planting cells in a 96-well plate, the total number of cells per well is required to be consistent (2000 / well), the amount of culture medium added should also be consistent, and a blank control (that is, the same volume of medium) is set, each well The final volume is 100ul. Place in a 37°C, 5% CO2 incubator and incubate for 24 hours.

[0072] (2) Add 10 ul of CCK-8 solution (purchased from Japan Dojin Chemical Research Institute, article number: CK04) to each well. The amount of CCK-8 solution can be increased or decreased according to specific conditions.

[0073] (3) Cultivate for 4 hours under normal culture conditions (37°C, 5% CO2 incubator) (th...

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Abstract

The invention discloses a new use of microRNA 145. The invention provides application of any matter in following 1) to 3) in preparation of products of facilitating migration, diffusion and / or invasion of tumour cells: 1) microRNA 145, 2) a recombinant vector containing encoding gene of the microRNA145, 3) recombinant virus containing encoding gene of the microRNA145, and 4) recombinant viral vector containing encoding gene of the microRNA145. Experimental results of the invention show that the microRNA145 cannot affect growth of HCT-8 cell after overexpression, but can obviously enhance invasion and migration ability of highly metastatic colorectal cancer cells, and play an important role in the transfer process of colorectal cancer, so that the microRNA 145 is related to colorectal cancer metastasis.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a new application of microRNA 145. Background technique [0002] Micro RNAs (miRNAs) are single-stranded small RNA molecules with a size of about 21-23 bases, which guide the silencing complex (RISC) to degrade mRNA or inhibit the translation of mRNA by base pairing with target gene mRNA, usually during transcription Post-level regulation of protein expression (recently found that micro RNA can also regulate gene expression at the transcriptional level). With the deepening of research on micro RNA, more and more evidences show that micro RNA molecules play the role of proto-oncogene or tumor suppressor gene in all aspects of tumor development. [0003] Michael et al. first used micro RNA cloning and Northern blot technology to detect the expression profile of micro RNAs in colorectal cancer and surrounding normal mucosal tissues, and found that miR-145 (micro RNA145, Geneban...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/63C12N7/01C12N15/86C12N5/10C12Q1/68A61K45/00A61P35/04
Inventor 马洁袁伟
Owner 普华赛尔生物医疗科技有限公司