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Application of RNASE4 serving as drug target to brain glioma inhibition drugs

A glioma and drug technology, applied in the application field of RNASE4 as a drug target in glioma inhibitory drugs, can solve unknown problems, achieve the effect of expanding the scope, overcoming non-selectivity, and improving prognosis

Inactive Publication Date: 2016-07-06
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether RNASE4 can be used as a therapeutic target for glioma and its mechanism of action are still unknown.

Method used

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  • Application of RNASE4 serving as drug target to brain glioma inhibition drugs
  • Application of RNASE4 serving as drug target to brain glioma inhibition drugs
  • Application of RNASE4 serving as drug target to brain glioma inhibition drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1, the expression of RNASE4 protein in human glioma tissue

[0060] In order to study the expression of RNASE4 gene in glioma tissues, 430 tissues of glioma patients with different malignant degrees were collected and a series of related experiments were done. First, the expression of RNASE4 protein in glioma was analyzed by conventional immunohistochemical method, the results are as follows figure 2 shown.

[0061] Nuclei are stained blue by hematoxylin, showing cell number distribution. The brown-yellow part of the tissue developed by DAB (diaminobenzidine) (that is, the part shown in dark black in the figure) represents the expression of RNASE4 protein.

[0062] The results showed that the stained positive tissue cells were arranged disorderly, the degree of compactness and looseness was inconsistent, and the shapes and sizes of the nuclei were different. They were overgrown tumor tissue cells, and the brownish yellow staining of the nuclei was also very r...

Embodiment 2

[0064] Example 2, siRNA or shRNA targeting the RNASE4 gene can inhibit the growth of glioma cells

[0065] The siRNA or shRNA targeting the RNASE4 gene described above was introduced into the glioma cell U87MG to silence the expression of the RNASE4 gene, thereby inhibiting the growth of the glioma cell. The transfection process can be mediated by either siRNA liposomes or lentiviruses containing shRNA.

[0066] Wherein the liposome transfection steps are as follows: inoculate 0.3x10 6 A glioma cell U87MG was transferred to a 35mm cell culture dish, and the DMEM cell culture solution of 2ml was added to each dish, and placed in an incubator for cultivation (cultivation conditions were: 5% CO 2 ; temperature is 37 degrees), overnight, after the cells adhere to the wall, when the cell adhesion rate reaches more than 60%, prepare the transfection reagent ( 2000 transfection reagent, purchased from American Life Company, the article number is 11668027) and siRNA (entrusted to S...

Embodiment 3

[0069] Example 3: shRNA targeting the RNASE4 gene inhibits the growth of xenografted tumors in nude mice

[0070] Cell lines stably expressing RNASE4 gene shRNA were used to determine the efficiency of gene silencing. Afterwards, we randomly divided 18 5-week-old male nude mice into three groups, and subcutaneously injected glioma U87MG stably infected cell lines (ControlshRNA cell line, RNase-4shRNA-1 cell line and RNase-4shRNA-2, these three lines The cell line was isolated from the above-mentioned cells infected with lentivirus), and each mouse was injected with 5x10 6 cells / 200 μL. The cell viability before and after the injection of the ControlshRNA cell line was 97.02% and 96.53%, the cell viability before and after the injection of the RNASE4shRNA-1 cell line was 98.35% and 97.69%, and the cell viability before and after the injection of the RNASE4shRNA-2 cell line was 96.15% and 93.29%, respectively. %, therefore, the subcutaneous injection procedure did not affect c...

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Abstract

The invention belongs to the field of biological medicine, relates to a novel brain glioma treatment target, in particular to ribonuclease 4 protein serving as the brain glioma treatment target, further relates to siRNA, shRNA and a neutralizing antibody of the ribonuclease 4 and includes application of the drug prepared from the siRNA, shRNA and the neutralizing antibody and serving as the target to inhibition of growth of brain glioma.The invention discloses application of RNASE4 serving as the drug target to brain glioma inhibition drugs, and the RNASE4 is the ribonuclease 4 and has the amino acid sequence as shown in SEQ ID NO: 1.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a new therapeutic target for glioma, specifically, it involves using ribonuclease 4 protein as a therapeutic target for brain glioma, and also relates to ribonuclease 4 siRNA, shRNA and The neutralizing antibody, including the medicine prepared by using the siRNA, shRNA and the neutralizing antibody, is used as an application for targeting and inhibiting the growth of brain glioma. Background technique [0002] Glioma generally refers to tumors of various neuroepithelial origins, and is the most common malignant tumor in the central nervous system, accounting for about 35% to 60% of all intracranial tumors. The term "glioma" covers a group of brain tumors including astrocytoma, oligodendroglioma, medulloblastoma, ependymoma. All gliomas grow in a malignant and infiltrating manner, and it is difficult to achieve pathological total resection even through surgical operations. Therefore, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/7105A61K39/395A61P35/00G01N33/68G01N33/574
CPCA61K31/7105A61K2039/505C07K16/40C07K2317/76G01N33/57407G01N33/57484G01N33/68
Inventor 盛静浩胡国富许正平陈光弟
Owner ZHEJIANG UNIV
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