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Application of miR-34a and target gene NOTCH1 to preparation of mesenchymal stem cell based glioma targeting carrier

A glioma-targeting, mesenchymal stem cell technology, applied in the application field of glioma-targeting vectors, can solve problems such as the unclear influence of mesenchymal stem cells on tumors

Inactive Publication Date: 2017-04-26
南京千年健干细胞基因工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Scott A. Bergfeld et al pointed out in the review article Bone marrow-derived mesenchymal stem cells and the tumor microenvironment that there are still serious differences in the scientific community on whether mesenchymal stem cells promote tumor growth or inhibit tumor production. The role of tumor type is not clear

Method used

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  • Application of miR-34a and target gene NOTCH1 to preparation of mesenchymal stem cell based glioma targeting carrier
  • Application of miR-34a and target gene NOTCH1 to preparation of mesenchymal stem cell based glioma targeting carrier
  • Application of miR-34a and target gene NOTCH1 to preparation of mesenchymal stem cell based glioma targeting carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: The effect of miR-34a targeting regulatory gene NOTCH1 on BMSC migration behavior

[0028] 1. Experimental materials

[0029] 1. miR-34a mimics and miR-34a inhibitor (purchased from Guangzhou Ribobio)

[0030] (1) miR-34a mimics double-stranded sequence:

[0031] Sense strand: 5'-UGGCAGUGUCUUAGCUGGUUGU-3'

[0032] Complementary strand: 5'-ACAACCAGCUAAGACACUGCCA-3'

[0033] (2) miR-34a mimics NC double-strand sequence:

[0034] Sense strand: 5'-UCACAACCUCCUAGAAAGAGUAGA-3'

[0035] Complementary strand: 5'-UCUACUCUUUCUAGGAGGUUGUGA-3'

[0036] (3) miR-34a inhibitor sequence:

[0037] 5'-ACAACCAGCUAAGACACUGCCA-3'

[0038](4) miR-34a inhibitor NC sequence:

[0039] 5'-UCUACUCUUUCUAGGAGGUUGUGA-3'

[0040] 2. miR-34a and U6qRT-PCR primer sequences (designed and synthesized by Shanghai Gemma)

[0041] miR-34a reverse transcription primer: 5'-GTCGTATCCAGTGCGTGTCGTGGAGTCGGCAATTGCACTGGATACGACAACAACC-3';

[0042] F: 5'-GGGTGGCAGTGTCTTAGC-3'; R: 5'-CAGTGCGTGTCG...

Embodiment 2

[0071] Example 2: Effects of modified BMSCs on the growth of glioma cells

[0072] The inventors studied the effect of miR-34a down-regulated bone marrow mesenchymal stem cells on the growth of glioma U87, glioma U251, and glioma C6, and found no promoting effect on the growth of glioma cells.

[0073]The present invention confirms that after the low expression of miR-34a in mesenchymal stem cells, the tropism of mesenchymal stem cells to glioma U87, glioma U251, and glioma C6 is all enhanced, and the targeting is increased; miR-34a is low After expression, the expression of NOTCH1 gene was up-regulated, and the content of NOTCH1 protein was significantly increased, which proves that miR-34a improves the tropism of mesenchymal stem cells to glioma U87, glioma U251, and glioma C6 by regulating its target gene NOTCH1. . Drugs that inhibit miR-34a and upregulate NOTCH1 gene and protein expression can be used to improve the targeting of mesenchymal stem cells to glioma, and the m...

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Abstract

The invention discloses application of miR-34a and target gene NOTCH1 to preparation of a mesenchymal stem cell based glioma targeting carrier. After miR-34a low expression of mesenchymal stem cells, cytotaxis of the mesenchymal stem cells to gliomas U87, gliomas U251 and gliomas C6 is enhanced completely, targeting performance is improved, and targeting specificity is achieved; after miR-34a low expression, gene NOTCH1 expression is up-regulated, and NOTCH1 protein content is increased evidently, so that miR-34a improves cytotaxis of the mesenchymal stem cells to gliomas U87, gliomas U251 and gliomas C6 by regulating the target gene NOTCH1. Medicines which inhibit miR-34a and up-regulate gene NOTCH1 and protein expression can be used for improving glioma targeting performance of the mesenchymal stem cells, and modified BMSC has no promotion effect on growth of glioma cells and can be made into the mesenchymal stem cell based glioma targeting carrier.

Description

technical field [0001] The invention belongs to the field of stem cells and relates to targeting tumor tissue using stem cells as a carrier, in particular to the application of miR-34a and its target gene NOTCH1 in preparing a mesenchymal stem cell-based glioma targeting carrier. Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells, MSCs) have the ability of self-renewal and multiple differentiation potentials. A large number of experiments have used this characteristic of MSCs to study new therapeutic methods for related diseases. Gene therapy is considered to be one of the most promising options for treating tumors after traditional therapies such as surgery, radiotherapy and chemotherapy. The primary problem of gene therapy is to choose a suitable carrier, and the tumorigenicity of mesenchymal stem cells makes it possible to become an effective carrier for tumor gene therapy. [0003] On the targeting effect of mesenchymal stem cells on tumors. S...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K48/00
CPCA61K48/0008C12N5/0663C12N2501/06
Inventor 袁春丽高云雷
Owner 南京千年健干细胞基因工程有限公司
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