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Application of a combination of miRNAs in the preparation of anti-non-small cell lung cancer drugs

A technology for non-small cell lung cancer and a composition, applied in the field of genetic engineering, can solve the problems of low survival rate, easy development of drug resistance in patients, loss of surgical opportunities, etc., and achieve the effect of promoting normalization

Inactive Publication Date: 2019-02-22
JIANGSU PROVINCE HOSPITAL
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  • Description
  • Claims
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Problems solved by technology

Non-small cell lung cancer accounts for 80% of all lung cancers, and more than two-thirds of lung cancer patients have metastasized at the time of diagnosis, often losing the opportunity for surgery, and the chemotherapy drugs for lung cancer are not effective, and patients are prone to drug resistance. The survival rate is less than 5%, and there is an urgent need to find new and effective treatments for lung cancer

Method used

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  • Application of a combination of miRNAs in the preparation of anti-non-small cell lung cancer drugs
  • Application of a combination of miRNAs in the preparation of anti-non-small cell lung cancer drugs
  • Application of a combination of miRNAs in the preparation of anti-non-small cell lung cancer drugs

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Experimental program
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preparation example Construction

[0033] The preparation method of pre-miR-1: 1. The pre-miR-1 sequence (pUC57-Simple-pre-miR-1) was synthesized by Nanjing Kaiji Biotechnology Development Co., Ltd. The target vector pYr-mir30-shRNA-2 was purchased from Changsha Yingrun Biotechnology Co., Ltd.; 2. The pre-miR-1 fragment was cloned into the pYr-mir30-shRNA-2 vector; 3. The clone was confirmed by sequencing; 4. Plasmid extraction.

[0034] The preparation method of pre-miR-206 is the same as that of pre-miR-1.

[0035] anti-miR-31 is an artificially synthesized miR-31 repressor that inhibits the expression of endogenous mature miR-31), and pre-miR-1, pre- The co-transfection of miR-206 and anti-miR-31 into A549 and H460 cells simultaneously up-regulated the expression of miR-1, miR-206 and down-regulated the expression of miR31.

Embodiment 1

[0037] 1. Differential expression of miRNAs in non-small cell lung cancer tumor-associated fibroblasts and normal fibroblasts

[0038] (1): Gene chip detection of miRNA expression levels in tumor-associated fibroblasts (CAF) and paired normal fibroblasts (NF) of 3 patients with non-small cell lung cancer;

[0039] 1. Chip preparation and analysis: According to the requirements of LC Sciences in the United States, lung cancer cell specimens were prepared and submitted to LC Sciences for chip preparation, and Sanger miRBase V10.0 version completed the chip analysis.

[0040] 2. Chip results: The experimental data comes from LC Sciences, and the results are as follows: figure 1 shown.

[0041] 3. Analysis of the results: Compared with NF, the down-regulation of miR-1 / miR-206 was the most obvious in CAF; the up-regulation of miR-31 was the most obvious.

[0042] (2) qRT-PCR analysis of the expression levels of miR-1, miR-206 and miR-31 in tumor-associated fibroblasts (CAF) and p...

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Abstract

The invention belongs to the field of genetic engineering, and particularly relates to application of a novel miRNA (micro-ribonucleic acid) composition to preparing non-small cell lung cancer resisting medicines. Transformation of lung tumor-associated fibroblasts of non-small cell lung cancer to normal fibroblasts can be promoted by means of changing expression level of the miRNA composition; the novel miRNA composition includes miR-1, miR-206 and miR-31. The application has the advantages that the miR-1 / miR-206 are simultaneously over expressed in the lung tumor-associated fibroblasts of the non-small cell lung cancer, the miR-31 is expressed at low level in the lung tumor-associated fibroblasts of the non-small cell lung cancer, accordingly, transformation of the lung tumor-associated fibroblasts to the normal fibroblasts can be promoted, anti-tumor effects can be realized at cell and animal level, and the miRNA composition has potential anti-tumor application value.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to the construction of a method for transforming non-small cell lung cancer tumor-associated fibroblasts into normal fibroblasts and its application in anti-tumor therapy Background technique [0002] Fibroblasts are key cellular components of human tissues and tumors and can be divided into resting and activated fibroblasts. For example, the fibroblasts are highly activated at wound healing sites. Activated fibroblasts invade the lesion and produce extracellular matrix (ECM), which acts as a scaffold for other cells. Once the wound is repaired, the activated fibroblasts return to a resting phenotype, which is called normal fibroblasts (Normal Fibroblasts, NFs). At present, it is believed that the development of tumor is not only determined by malignant cancer cells, but also requires the participation of activated tumor fibroblasts or tumor-associated fibroblasts (Carci-noma-as...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K31/7088A61P35/00
CPCA61K31/7088A61K2300/00
Inventor 沈华束永前刘连科
Owner JIANGSU PROVINCE HOSPITAL
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