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Polypeptide with function of inhibiting tumor cell proliferation and application

A technology for the proliferation and function of tumor cells, which is applied to polypeptides and application fields that can inhibit tumor cell proliferation, can solve the problems of large overall molecular weight, and achieve the effects of good tissue permeability, low immunogenicity, and small molecular weight

Active Publication Date: 2021-03-16
ZHEJIANG MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Mu4 in this application has a total of 572 amino acids, and the overall molecular weight is relatively large, which needs further improvement

Method used

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  • Polypeptide with function of inhibiting tumor cell proliferation and application
  • Polypeptide with function of inhibiting tumor cell proliferation and application
  • Polypeptide with function of inhibiting tumor cell proliferation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Construction of Ku70-C polypeptide expression vector pBICEP-N-Flag-Ku70-C targeting Skp2

[0017] (1) Preparation of pBICEP-N-Flag-Ku70-C and pBICEP-N-Flag-Ku70-Mu-C expression vectors

[0018] By mutation PCR method, from the wild-type full-length Ku70 expression vector pBICEP-N-Flag-Ku70 (see publication number CN111333712A for the full sequence of the gene of the Ku70 protein gene GenBank No. NM_001288977) to amplify and express the Ku70-C polypeptide ( The amino acid sequence is shown in SEQ ID No.1, the encoding gene sequence is shown in SEQ ID No.2) pBICEP-N-Flag-Ku70-C expression vector, and the control vector pBICEP-N-Flag-Ku70-Mu-C The control vector pBICEP-N-Flag-Ku70-Mu-C expresses the mutant Ku70 protein with missing C-terminal (the mutant Ku70 protein with missing C-terminal is represented by Ku70-Mu-C, and the amino acid sequence is shown in SEQ ID No.3 shown, the coding gene sequence is shown in SEQ ID No.4).

[0019] The specific method for ...

Embodiment 2

[0055] Example 2: Detection of the inhibitory activity of Ku70-C polypeptide on the proliferation of lung cancer cell lines A549, NCI-H1975 and cervical cancer cells HeLa

[0056] (1) Culture lung cancer cell lines A549, NCI-H1975, cervical cancer cell HeLa and colorectal cancer cell HCT-116;

[0057] The cell culture medium is the complete medium including RPMI-1640 medium (A549, NCI-H1975, HCT-116) and complete MDEM medium (HeLa). The culture conditions are as follows: each cell is seeded at a density of 1×10 per well. 5 Cells were seeded in a 96-well plate at a culture temperature of 37°C until 50%-60% confluence.

[0058] (2) Lung cancer cell lines A549, NCI-H1975, cervical cancer cells HeLa and colorectal cancer cells HCT-116 were transferred into pBICEP-N-Flag-Ku70-C, pBICEP-N-Flag-Ku70-C and pBICEP-N-Flag- Ku70-Mu-C and pBICEP-N-Flag-Ku70 plasmids;

[0059] A549, NCI-H1975, HeLa and HCT-116 cells were cultured in 96-well plates to 50%-60% confluency for liposome tran...

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PUM

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Abstract

The invention discloses a polypeptide with a function of inhibiting tumor cell proliferation and application. The polypeptide is a Ku70 protein truncation, and the amino acid sequence of the polypeptide is shown as SEQ ID No. 1. According to the polypeptide, the polypeptide Ku70-C capable of effectively inhibiting tumor proliferation is separated from Ku70 protein by using a mutation PCR method. In-vitro anti-tumor activity detection shows that the modified protein shows good anti-tumor proliferation characteristics and can be used for optimizing anti-tumor polypeptide, and a model is providedfor developing anti-tumor drugs. The polypeptide Ku70-C only has 74 amino acid residues and is relatively small in molecular weight, low in immunogenicity and good in tissue permeability. Certain application prospects are realized in the field of medicine.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a polypeptide with the function of inhibiting tumor cell proliferation and its application. Background technique [0002] Traditional tumor treatment methods mostly use surgery, radiation or chemical drug treatment, which have large side effects and are prone to recurrence. Even the emerging tumor immunotherapy still cannot overcome the shortcomings of tumor treatment drugs such as large side effects and easy recurrence. Finding safe and effective tumor-targeted therapy drugs is an urgent problem in the field of tumor treatment. [0003] Peptide drugs have low immunogenicity, are easy to synthesize and transform, and have good tissue permeability. They have shown good application prospects in the treatment of tumors, cardiovascular diseases, immune diseases and other diseases. At present, there are more than 80 kinds of peptide drugs on the market in the world, and nearly 20...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12A61K48/00A61K38/17A61P35/00
CPCC07K14/47A61K48/005A61P35/00A61K38/00
Inventor 贾静杨新燕晏沙沙王屹喆
Owner ZHEJIANG MEDICAL COLLEGE
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