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Short peptide for inhibiting tumors as well as coding gene and application of short peptide

A technology that encodes genes and tumors. It is applied in applications, anti-tumor drugs, and genetic engineering. It can solve problems such as structural instability, long peptide chains, and difficult chemical synthesis.

Active Publication Date: 2017-08-29
SHANTOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This kind of anti-tumor peptide directly derived from the body generally has a long peptide chain, which is difficult to synthesize by chemical methods, difficult to purify, and has the disadvantages of unstable structure and easy inactivation.

Method used

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  • Short peptide for inhibiting tumors as well as coding gene and application of short peptide
  • Short peptide for inhibiting tumors as well as coding gene and application of short peptide
  • Short peptide for inhibiting tumors as well as coding gene and application of short peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Solid-phase synthesis, cleavage, purification and identification of tumor-inhibiting short peptides

[0027] The present invention utilizes a polypeptide synthesizer to synthesize the short peptide for inhibiting tumors by a solid-phase synthesis method:

[0028] 1) With DMF as the solvent, the concentration of various α-amino acids protected by Fmoc is 0.25M, the concentration of HBTU solution and HOBt solution is 0.33M, the concentration of piperidine solution is 200ml / L, and the concentration of DIEA solution is 174.2ml / L.

[0029] 2) Weigh 0.05mmol of Fmoc-Ala-Wang resin (functional group content 0.33mmol / g) into a solid-phase reactor, add 8ml of DCM to swell overnight, and remove the solvent under reduced pressure. Add 8ml of piperidine solution with a concentration of 200ml / L, react at room temperature for 5min, and drain; then add 8ml of piperidine solution with a concentration of 200ml / L, react for 20min at room temperature, and drain; wash with 8ml ...

Embodiment 2

[0038] Example 2: Determination of antitumor activity of short peptides that inhibit tumors

[0039] (1) Use human cervical cancer cells (HeLa) in logarithmic phase of growth, digest and resuspend the cells to obtain a cell suspension, and dilute the cells to 50,000 cells / mL. Add 100 µL to each well of the 96-well plate, put the paved 96-well plate in the incubator, and store at 37°C, 5% CO 2 Incubate overnight (>8 hours) under conditions.

[0040] (2) Use 0.01M pH7.4 PBS to dissolve the short peptide at 1 mg / mL. Dilute the short peptide sample to 50 µg / mL with complete medium. Discard the cell culture medium cultivated overnight, add 100 µL of 50 µg / mL polypeptide to each well, and repeat 3 times in 4 parallels to each well. At the same time, an equal volume of 50 μg / ml 5-FU and 0.01M pH7.4 PBS were used as positive and negative controls, at 5%37 o C conditions were cultured for 18 h.

[0041] (3) Next, discard the culture medium and dry it, add 50 μL of ice-cold 500 g / L...

Embodiment 3

[0046] Example 3: Comparison of the antitumor activity of short peptides that inhibit tumors and other short peptide fragments in hemocyanin

[0047] (1) Synthesize 12 different hemocyanin short peptide fragments and name them respectively (B1, B2, B3, B9, B10, B13, B14, S7, S8, S9, S10). The short peptides of the present invention are named B11 .

[0048] (2) Use human cervical cancer cells (HeLa) in the logarithmic phase of growth, digest and resuspend the cells to obtain a cell suspension, and dilute the cells to 50,000 cells / mL. Add 100 µL to each well of the 96-well plate, put the paved 96-well plate in the incubator, and store at 37°C, 5% CO 2 Incubate overnight (>8 hours) under conditions.

[0049] (3) Use 0.01M pH7.4 PBS to dissolve the short peptide at 1mg / mL. Dilute different short peptide samples to 50 µg / mL with complete medium. Discard the cell culture medium cultivated overnight, add 100 µL of 50 µg / mL polypeptide to each well, and repeat 3 times in 4 paralle...

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Abstract

The invention discloses a short peptide for inhibiting growth of tumor cells as well as a coding gene and application of the short peptide. The amino acid sequence of the short peptide is represented by SEQ ID NO:1. The short peptide provided by the invention is capable of inhibiting the growth of cervix uterus cells (HeLa) and can be a good anti-cancer drug with good low side effects.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a tumor-suppressing short peptide, its coding gene and its application. Background technique [0002] Cancer has high morbidity and mortality rates worldwide. In recent decades, human beings have made unremitting efforts to treat cancer, and the research on cancer mainly focuses on developing new treatment methods and reducing the side effects of treatment methods on patients. The commonly used cancer treatment methods are mainly surgery or chemotherapy, but in many cases not only cannot treat patients efficiently, but also have a high risk of complications. Some cancer treatment drugs not only have multiple side effects, but also may cause cause multidrug resistance. [0003] As a new type of drug for treating cancer, antitumor peptides are attracting more and more attention because they are selective for tumor cells and not easy to cause drug resistance. [0004] Many antitumor ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/795C12N15/12A61K38/41A61P35/00
CPCA61K38/00C07K14/795
Inventor 章跃陵刘尚杰黄河郑莉媛王帆
Owner SHANTOU UNIV
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