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Antitumor recombinant protein GPTI, coding gene and applications thereof

A recombinant protein, anti-tumor technology, applied in the field of bioengineering, can solve the problems of inability to kill tumor cells, reduced blood picture of patients, bleeding, etc., and achieve the effects of high anti-tumor activity, simple operation, and small toxic and side effects.

Inactive Publication Date: 2017-10-13
GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently used chemotherapeutic drugs and radiotherapy can damage all active proliferating cells in the body, such as the hematopoietic system and the gastrointestinal tract, so patients undergoing radiotherapy and chemotherapy often have decreased blood counts, gastrointestinal bleeding, nausea, and vomiting and other symptoms
Moreover, due to the limitations of the patient's physical and mental endurance and drug resistance of tumor cells, radiotherapy and chemotherapy cannot kill all tumor cells, leaving hidden dangers for tumor recurrence and metastasis and disease progression.
Biological therapy and traditional Chinese medicine therapy are also reasonable methods of anti-tumor, but still need to continue research to improve the efficacy

Method used

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  • Antitumor recombinant protein GPTI, coding gene and applications thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Escherichia coli in vitro expression of anti-tumor recombinant protein:

[0053] 1. Synthesis of artificial gene: artificially synthesize the partial or full-length sequence of SEQ ID No.3 (the gene encoding cardiac troponin I) in the sequence table, and add BamH I and 3' end to its 5' end. NotI restriction site, to obtain the cloned sequence of SEQ ID No.3 in the sequence listing. Design and synthesize part or all of the nucleic acid sequence of the gene encoding the amino acid residues of the artificial short peptide of SEQ ID No.2 in the sequence listing, and add NdeI and BamH I restriction sites at its 5' end and 3' end to obtain the sequence The cloned sequence of SEQ ID No.4 in the list. The nucleic acid sequence of the gene encoding the amino acid residues of the artificial short peptide and the nucleic acid sequence of the gene encoding the amino acid residues of cardiac troponin I were respectively digested with BamH I, purified and connected, and the nucleic ...

Embodiment 2

[0063] Effects of the anti-tumor recombinant protein of the present invention on the proliferation function of cultured vascular endothelial cells:

[0064] Human umbilical vein endothelial cell EVC304 cell line was purchased from PLA Academy of Military Medical Sciences; RPMI-1640 medium, fetal bovine serum, and trypsin were purchased from Hyclone Company, and other reagents were purchased from Sigma Company, and the anti-tumor recombinant protein (GPTI1) was purchased from 89% expressed the purified product.

[0065] ECV304 cells were cultured according to the conventional method of endothelial cells. The cultured ECV304 cells with normal growth were inoculated into 96-well plates at 5*104 cells per well, cultured at 37°C for 12 hours, and then replaced with purified anti-tumor recombinants containing the expression of the sequence GPTI 1 Protein 5 μg / ml fresh culture solution, the blank control was replaced with fresh culture solution containing an equal volume of normal sali...

Embodiment 3

[0067] In vivo tumor suppression experiment of anti-tumor recombinant protein of the present invention: Take 24 Kunming mice and divide them into phosphate buffer group and expression purification group, inoculate the neck with S180 ascites cell tumor strain subcutaneously, and inject phosphoric acid subcutaneously three days later. Salt buffer solution and expression purification solution 1.0 mg protein / kg were injected every 12 hours for 7 consecutive days. The tumor tissues were observed, cut out, and weighed. The results showed that the tumor weights in the phosphate buffered saline and expression-purified liquid sequence GPTI groups were 1.11±0.26 grams and 0.16±0.07 grams, respectively, and the tumor inhibition rate was 85.6%.

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Abstract

The invention discloses an antitumor recombinant protein GPTI, a coding gene and applications thereof, and provides an antitumor recombinant protein and a coding gene thereof, and a method for efficiently expressing the protein, wherein the recombinant protein GPTI is the protein formed by partially and completely fusing the amino acid residue at the amino terminal of cardiac troponin I and the amino acid residue sequence at the carboxy terminal of an artificial short peptide having the amino acid residue sequence represented by SEQ ID No.2. According to the present invention, the antitumor recombinant protein GPTI as the drug can be clinically used for treating a variety of solid malignant tumors, has characteristics of high efficiency, broad spectrum, low toxic-side effect and the like, and can be expected to be the completely-new antitumor drug.

Description

technical field [0001] The invention relates to the field of bioengineering. More specifically, the present invention relates to an anti-tumor recombinant protein GPTI and its coding gene and application. Background technique [0002] Malignant tumors are the most serious diseases that endanger human health. The common methods for the treatment of cancer at present are: surgical operation, chemical drug therapy, radiation therapy, biological therapy and traditional Chinese medicine therapy. Among them, surgery, chemotherapy, and radiotherapy are the most commonly used. Surgical treatment, as the first choice and main method of most tumor treatment, has made countless tumor patients recover, prolong life or improve the quality of life. However, in terms of tumor treatment, surgical treatment is only suitable for patients with certain conditions and ranges, and due to the possibility of tumor recurrence and metastasis, surgical resection does not always achieve the purpose ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N1/21A61P35/00
CPCC07K14/00C07K14/47C07K2319/00
Inventor 刘凤鸣师凤华谭木秀
Owner GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS
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