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Gene medicine for promoting differentiation of tumor stem cells and applications thereof

A technology of tumor stem cells and gene medicine, which is applied in the field of gene medicine that promotes the differentiation of tumor stem cells, can solve the problems of recurrence and metastasis that plague surgeons, produce tumor tissue treatment resistance, and lack of differentiation of tumor stem cells, so as to improve drug sensitivity, Safe to use, low toxicity and side effects

Inactive Publication Date: 2015-04-08
SHANGHAI PULMONARY HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Traditional tumor treatment methods include chemotherapy, radiotherapy and surgery, but from the perspective of clinical treatment effects, none of them can completely cure the tumor
Moreover, due to the non-specific killing effect of radiotherapy and chemotherapy, the toxicity to the body is relatively high, and long-term use will cause treatment tolerance of tumor tissues.
The curative effect of surgery on patients with early tumors is obvious, but many tumors are found late and cannot be implemented, and the problem of postoperative recurrence and metastasis has always troubled surgeons
However, since Nell-1 is an osteogenic gene, most of its research at home and abroad focuses on its bone differentiation-related functions, and there is no research on Nell-1 promoting the differentiation of tumor stem cells.

Method used

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  • Gene medicine for promoting differentiation of tumor stem cells and applications thereof
  • Gene medicine for promoting differentiation of tumor stem cells and applications thereof
  • Gene medicine for promoting differentiation of tumor stem cells and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Application of Nell-1 gene to glioma

[0025] Use the RT-PCR method to obtain the Nell-1 gene sequence. After the test is correct, the full length of the gene is amplified by the PCR method in vitro, and then the amplified product is cloned into the expression plasmid to obtain the expression plasmid carrying the Nell-1 gene (Nell-1 gene). 1 expression plasmid). The Nell-1 expression plasmid was used to transfect into a glioma stem cell line (DA66), and the positive ratio of CD133 in the tumor stem cell line was about 65-70%.

[0026] After screening by G418, a cell line (DA66-Nell-1) with stable and high expression of Nell-1 was obtained, stained with CD133 antibody, and then analyzed by flow cytometry, such as figure 1 As shown (where DA66 is a complete blank control without any treatment, and DA66-Neo is a blank plasmid transfection), Nell-1 overexpression significantly reduces the expression of CD133, while blank plasmid transfection (DA66-Neo) has no eff...

Embodiment 2

[0031] Embodiment 2 object gene is the preparation of the recombinant of Nell-1

[0032] Use the RT-PCR method to obtain the Nell-1 gene sequence. After the test is correct, the full length of the gene is amplified by PCR in vitro, and the amplified product clone and the adenovirus shuttle plasmid are cut with a specific endonuclease to construct a Nell-1-carrying gene. -1 gene shuttle plasmid, then subcloned into pQB1-AdCMV5 (containing adenovirus E1A), and co-transfected with QB1-viralDNA, positive virus clones were screened by PCR to obtain Ad-Nell-1 recombinant, amplified, transfected HEK293 cells were transfected, cultured, crudely separated Nell-1 gene recombinant adenovirus, purified by ultracentrifugation, and sterilized.

Embodiment 3

[0033] The preparation of embodiment 3 injections

[0034] The Ad-Nell-1 recombinant 2 * 10 that embodiment 2 is made 15 For virus titer, add physiological saline for injection to 1000mL, filter aseptically, and dispense into 2mL vials.

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Abstract

The invention provides a gene medicine for promoting the differentiation of tumor stem cells. The gene medicine comprises a recombinant of which the target gene is a Nell-1 gene and can effectively induce the differentiation of the tumor stem cells, inhibit the multiplication of tumor cells, reduce the drug resistance of the tumor cells and improve the drug sensitivity and can be combined with chemical treatment to enhance the chemical treatment effect; and the gene medicine is low in toxic and side effects and safely used. A carrier of the recombinant can be an expression plasmid or a virus carrier, preferably the virus carrier; the virus carrier is preferably selected from one of an adenovirus carrier, a herpes simplex virus carrier, a retroviruse carrier, an adeno associated virus carrier and a slow virus carrier; and most preferably, the virus carrier is the adenovirus carrier.

Description

technical field [0001] The invention relates to a gene drug for treating tumors, in particular to a gene drug for promoting tumor stem cell differentiation and its application. Background technique [0002] Traditional tumor treatment methods include chemotherapy, radiotherapy and surgery, but from the perspective of clinical treatment effects, none of them can completely cure the tumor. Moreover, due to the non-specific killing effect of radiotherapy and chemotherapy, the toxicity to the body is relatively high, and long-term use will cause treatment tolerance of tumor tissues. Surgery has obvious curative effect on patients with early tumors, but many tumors are found late and cannot be implemented, and the problem of postoperative recurrence and metastasis has always troubled surgeons. If the difference in gene expression between tumor cells and normal cells is grasped, the method of virus-mediated transfection can be used to adjust the expression of genes in tumor cells...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P35/00
Inventor 刘根桃蒋欣泉
Owner SHANGHAI PULMONARY HOSPITAL
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