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Application of 183aa micro-molecule polypeptides

A small molecule polypeptide, 1.183aa technology, applied in the application field of 183aa small molecule polypeptide, to achieve the effect of promoting cell apoptosis

Active Publication Date: 2016-09-07
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, despite the use of novel chemotherapeutic agents and improved treatment options, drug resistance remains a major problem in the treatment of multiple myeloma

Method used

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  • Application of 183aa micro-molecule polypeptides
  • Application of 183aa micro-molecule polypeptides
  • Application of 183aa micro-molecule polypeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1183aa

[0016] Example 1 183aa polypeptide recombination

[0017] 1) Cloning vector construction of recombinant peptide: extract human mRNA, reverse transcribe it into cDNA, use cDNA as template, upstream primer is 5'-GGGGTACCATGCACCTGCTCCCTCTCTGG-3' (containing KPN I restriction site); downstream primer is 5'- CGGGATCCTACTGTGTTCATCATACTGTCGAA-3' (with Bamh I restriction site).

[0018] The 552bp mRNA sequence corresponding to 183aa was successfully amplified by PCR technology, as shown in SEQ ID NO.2, and the amino acid sequence of the expressed 183aa small molecule polypeptide was shown in SEQ ID NO.1. The amplified fragment was ligated with the p3×FLAG-CMV-14 vector ( figure 1 ), transfer the ligated product into competent Escherichia coli DH5α, select clones on the Amp-containing agar plate, extract the recombinant plasmid with alkaline lysis method, and identify it with KPN I and Bamh I enzymes.

[0019] The main process of the above operation is as follows:

[0020] The STAT1...

Embodiment 2

[0028] Example 2 Detection of Anti-Multiple Myeloma Function of Recombinant Peptides

[0029] 1) CCK-8 experiment proves the anti-proliferation effect of recombinant peptides on multiple myeloma cells at the cellular level

[0030] CCK-8 cell proliferation experiment: In RPMI 8226 cells, the cells were collected 48 hours after transfection with wild-type and 183aa fragment truncated STAT1 vectors, and 5000 cells per well were seeded into 96-well plates with a volume of 100 μL per well. Change the medium in each well (the ratio of CCK-8 to culture medium is 1:10), incubate at 37°C for 2 hours, change the medium after reaching the time point, add 100 μL of cell culture medium containing 10 μL of CCK-8 solution to each well (be careful not to Air bubbles are formed in the well and they will affect the O.D reading). Incubate the culture plate in the incubator for 1.5-2h. (The same time should be kept at different time points) Measure the absorbance at (measurement wavelength 490...

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Abstract

The invention discloses application of 183aa micro-molecule polypeptides to preparation of anti-cancer drugs. By means of the bioengineering technology, a section of DNA sequence corresponding to 183 amino acid polypeptides is genetically recombined to a p3*FLAG-CMV-14 eukaryotic expression vector. Digestion and sequence analysis prove that after recombination is successful, the recombined polypeptides are transfected to multiple myeloma RPMI8226 cells, the protein expression of the polypeptides is detected through western blot, and recombination of the polypeptides is achieved. Then, studying is carried out on the function of the polypeptides to resist multiple myeloma, and cytology experiments show that the polypeptides have an important anti-cancer function of promoting myeloma cell apoptosis. An experiment basis is provided for new target spot developing on oncotherapy, and the 183aa micro-molecule polypeptides have very great development and application prospects when applied to tumorous clinic treatment.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the application of 183aa small molecule polypeptide. Background technique [0002] Multiple myeloma is an incurable malignant plasma cell disease with an incidence of 1-2% of tumors and 10-15% of hematologic malignancies. To date, multiple myeloma remains the second most common hematological malignancy. Despite improvements in conventional treatments, multiple myeloma remains incurable, with a 5-year survival rate of only 31%. Currently, chemotherapy is one of the most effective treatments for multiple myeloma. However, despite the use of novel chemotherapeutic agents and improved treatment options, drug resistance remains a major problem in the treatment of multiple myeloma. [0003] Peptide drugs are currently the most growing new field in the field of biomedicine. With their high efficiency, safety, and strong specificity, they are gradually used in the prevention an...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61P35/00C12N15/11C12N15/85
CPCA61K38/16C07K14/00
Inventor 程纯沈爱国王燏婵钟菲
Owner NANTONG UNIVERSITY
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