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Virus construct for treating liver cancer as well as application and construction method thereof

A technology for the treatment of liver cancer, constructs, applied in the field of biotechnology and gene therapy, can solve problems such as high therapeutic index

Inactive Publication Date: 2020-11-10
药鼎(北京)国际细胞医学技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These features lead to a high therapeutic index

Method used

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  • Virus construct for treating liver cancer as well as application and construction method thereof
  • Virus construct for treating liver cancer as well as application and construction method thereof
  • Virus construct for treating liver cancer as well as application and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 - Construction of viral constructs

[0078] In the present invention, restriction endonucleases HindⅢ / SalⅠ / EcoRI / XbaⅠ were purchased from New England Biolabs; PCR amplification reagents were purchased from TARAKA Baori Medical Biotechnology (Beijing) Co., Ltd.; endotoxin-free plasmid extraction kit DP117, DP118 was purchased from Tiangen (TIANGN) Biochemical Technology (Beijing) Co., Ltd.; pBHGlox_E1,3Cre recombinant adenovirus backbone vector and pDC316 adenovirus vector were purchased from Changsha Youbao Biology; GMSF-Flag and AFP fragments and all amplification primers were in Synthesized by Sangon Biotech (Shanghai). DNA recovery kit was purchased from Promega.

[0079] The invention discloses a method for constructing a novel oncolytic virus for individualized targeted therapy of liver cancer, which comprises the following steps:

[0080] Step 1: Construction of Ade-PDC316-CMV-GFP plasmid: In order to facilitate the detection of the expression of vario...

Embodiment 2

[0101] Example 2—oncolytic effect verification experiment

[0102] In the present invention, the novel oncolytic virus Ade AFP+RGD-4C+GMSF-Flag It has a selective inhibitory effect on liver cancer cells, does not infect normal liver cells, and has a very low infection rate in other tumor cells.

[0103] 1.MTT experiment

[0104] MTT is a good method to reflect the viability of cells. The present invention uses MTT to detect (0,0.001,0.01,0.1,1Ade AFP+RGD-4C+GMSF-Flag The effect on the viability of liver cancer cell HepG2 and normal cell HL7702, found that this new virus Ade AFP+RGD-4C+GMSF-Flag Can significantly inhibit the viability of liver cancer cells HepG2, but has no effect on the viability of normal liver cells HL7702 (see figure 2 )

[0105] 2. Plate colony formation experiment

[0106] Clonogenicity is a unique property of malignant cells. Plate colony formation assay can well reflect the colony formation ability of tumor cells. The present invention detects ...

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Abstract

The invention relates to a virus construct for treating liver cancer as well as application and a construction method of the virus construct. The invention also relates to a novel oncolytic virus forindividualized targeted therapy of liver cancer and a construction method thereof. The construct of the invention comprises an AFP promoter, an RGD-4C gene and a GMSF-flag gene, and optionally a glutamyl transpeptidase gene and a tumor-associated antigen gene, in an operably linked manner.

Description

technical field [0001] The invention relates to the fields of biotechnology and gene therapy. Specifically, the present invention relates to a virus construct for treating liver cancer, its use and construction method. The invention also relates to a novel oncolytic virus for individualized targeted therapy of liver cancer and its construction method. Background technique [0002] Liver cancer refers to malignant tumors of the liver and is generally divided into two categories: primary liver cancer and secondary liver cancer. Primary liver cancer originates from the epithelial or mesenchymal tissue of the liver, and is a high-incidence cancer in my country. Compared with primary liver cancer, secondary or metastatic liver cancer mainly refers to cancers that have metastasized to various organs of the body and invade the liver. There are many reasons for inducing liver cancer, such as hepatitis B and C virus infection, drinking water pollution, chemical substances causing c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/861C12N15/66C12N15/27C12N15/54C12N7/01A61K35/761A61P35/00A61P1/16
CPCA61K35/761A61P1/16A61P35/00C07K14/535C12N7/00C12N9/1044C12N15/66C12N15/86C12N2710/10332C12N2710/10343C12N2710/10352C12N2800/107C12N2830/008C12Y203/02013
Inventor 宫晓艳秦苗苗刘进稳周海涛
Owner 药鼎(北京)国际细胞医学技术有限公司
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