Anti-oncogene magnetic composite nanoparticle and preparation method thereof

Abstract

The invention relates to an anti-oncogene magnetic composite nanoparticle and a preparation method thereof. Egr1 and Hsp70 promoters and a HSVTK gene are obtained through PCR amplification and are connected to a pCDNA3.1 plasmid through the steps of enzyme digestion and connection, a magnetic nanoparticle is used as a gene carrier to guide the constructed gene into an HEK293 cell, and the expression of the HSV-TK gene is quantitatively observed after the induction of radiation or magnetic hyperthermia or the combination of radiation and magnetic hyperthermia. The expression quantity of the target gene is the highest after the Egr1 promoter of the plasmid is induced by 2GyX rays. The Egr1 and Hsp70 promoters both have high inducible expression activities, the target gene expression is induced by radiation or magnetic hyperthermia or the combination of radiation and magnetic hyperthermia, the capability of inducible expression on the gene is better in the combined effect of the radiation and magnetic hyperthermia, and an experimental basis is provided for the further comprehensive treatment of cancers.

Description

technical field [0001] The invention relates to the construction of a double-promoter pCDNA3.1-Egr1-Hsp70-HSVTK gene induced by radiation and heat shock for the treatment of cancer, magnetic composite nanoparticles and preparation thereof. Background technique [0002] In recent years, many new gene therapy methods (including radiation-gene therapy and gene-hyperthermia therapy) have been successively used in the treatment of liver cancer. Among them, the target gene with more applications and relatively positive curative effect is the HSV-TK / GCV system. The target gene it uses is a suicide gene or a drug-sensitive gene. Its therapeutic principle is to convert non-toxic drug precursors (such as GCV, etc.) into toxic drugs. It disrupts the synthesis of cellular DNA, thereby selectively killing rapidly proliferating tumor cells. However, it is very difficult to achieve a comprehensive tumor gene therapy and obtain an ideal curative effect until the problem of gene delivery is...

AI Technical Summary

Problems solved by technology

The radiation-gene therapy research using the radiosensitivity of the Egr-1 gene promoter regulatory sequence has achieved encouraging results, but this method still faces the specificity, safety, and gene expression efficiency in gene therapy. The low time is short, the cytotoxicity is small, and the gene expression after transfer lacks effective regulation methods, etc.

Moreover, the hypoxic environment commonly found in solid tumors can also reduce the induction activity of radiation promoters, which affects the efficacy and application of this therapy.

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