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Short chain polypeptide for inhibiting cancer cell growth as well as encoding gene and application of short chain polypeptide

A technique for encoding genes and tumor cells, applied to short peptides that inhibit the growth of cancer cells and their encoded genes and application fields, which can solve problems such as loss of inhibiting cell growth.

Active Publication Date: 2014-09-17
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Type II tumor suppressor gene is a novel tumor suppressor gene. They have a certain level of expression in normal cells and have certain functions, but in tumor cells, the DNA does not change, and the transcription and translation of the genes are different. Down-regulated to a certain extent, thus losing the effect of inhibiting cell growth

Method used

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  • Short chain polypeptide for inhibiting cancer cell growth as well as encoding gene and application of short chain polypeptide
  • Short chain polypeptide for inhibiting cancer cell growth as well as encoding gene and application of short chain polypeptide
  • Short chain polypeptide for inhibiting cancer cell growth as well as encoding gene and application of short chain polypeptide

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Experimental program
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Effect test

Embodiment 1

[0042] Embodiment 1, the acquisition of short peptide

[0043] The short peptide sequence used in the present invention is from a second human tumor suppressor, and its DNA fragment can be obtained by PCR amplification using a human liver cDNA library as a template.

[0044] Using the human liver cDNA library as a template and using h2cm7-N / h2cm7-C as primers, perform PCR amplification to obtain the PCR product of the short peptide gene fragment, which has BspEI and XbaI restriction sites at both ends.

[0045] h2cm7-N: 5'-CGATTCTCCGGACGCAGTGACCAGGTCAGAGAT-3' (SEQ ID NO: 4);

[0046] h2cm7-C: 5'-CTAGTCTAGATTATTGCTTTTGTCGCTTGTTTC-3' (SEQ ID NO: 5).

[0047] The specific PCR reaction system is as follows:

[0048]

[0049] Reaction conditions:

[0050] 94℃ 5min

[0051] 2 94°C 40sec

[0052] 3 60℃ 40sec

[0053] 4 72℃ 45min

[0054] 72°C 30min

[0055] 4°C 10min

[0056] 2, 3, 4 need 30cycles for cycle reaction

[0057]A 136bp PCR product was obtained...

Embodiment 2

[0060] Example 2, Functional Identification of Short Peptide REVC37

[0061] 1. Construction of recombinant vector containing short peptide REVC37 gene sequence

[0062] 1. Construction of fusion Tag BFP fluorescent protein vector pcDNA3.1-BFPC

[0063] 1) Preparation of Tag BFP gene fragment

[0064] Tag BFP is synthesized from the whole gene by Bomed Technology Development Co., Ltd. according to the sequence of Gene Bank. The Gene Bank number of Tag BFP is ABP88744.1 (the nucleotide sequence is sequence 3).

[0065] Use the plasmid containing the Tag BFP sequence obtained by whole gene synthesis as a template (or artificially synthesized sequence 3 as a template), and use BFP-N / BFP-C as primers to perform PCR amplification, and the PCR product obtained is the Tag BFP gene fragment , the two ends of the gene fragment have NotI and ClaI restriction sites respectively.

[0066] BFP-N: 5'-TAATGCGGCCGCATGAGCGAGCTGATTAAGGAG-3'

[0067] BFP-C: 5'-ACATATCGATATTAAGCTTGTGCCCCAGTT...

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Abstract

The invention discloses short chain polypeptide for inhibiting cancer cell growth as well as encoding gene and application of the short chain polypeptide. The polypeptide provided by the invention is the following polypeptide (a) or (b), wherein (a) is the polypeptide formed by amino acid sequences shown as a sequence 2 in a sequence table, (b) is the polypeptide derived from the (a), and the polypeptide (b) is obtained through the substitution and / or deletion and / or addition of one or a plurality of amino acid residues on the amino acid residue sequence of the sequence 2 in the sequence table and has the functions of inhibiting the tumor cell growth and / or promoting the tumor cell apoptosis. Experiments prove that the polypeptide provided by the invention can cause the apoptosis of HeLa cells of human cervical cancer cells, so the short chain polypeptide can become good anti-cancer medicine with low side effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a short peptide for inhibiting the growth of cancer cells, its coding gene and its application. Background technique [0002] Cancer, also known as malignant tumor, is a disease caused by the abnormal precise regulation mechanism of cell proliferation and apoptosis. In addition to uncontrolled growth, cancer cells will also locally invade the surrounding normal tissues and even spread or transfer to other parts of the body through the circulatory system in the body. organ. Cancer has become one of the leading killers of human health in the world. According to the announcement issued by the World Health Organization (WHO) in 2010, 12 million people are diagnosed with cancer and 7.6 million people die of cancer every year. If left unchecked, it is estimated that by 2030, the annual number of cancer deaths may reach 17 million, and most of them will occur in low- and middle-income deve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/12C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A61K38/17A61K48/00A61P35/00
Inventor 夏斌林坚任晓白魏贺佳
Owner PEKING UNIV
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