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Polypeptide derived from mammary gland adipose tissue and anti-tumor application thereof

An adipose tissue-derived technology, applied in the field of anti-tumor, can solve the problem of lack of available therapeutic targets for triple-negative breast cancer

Active Publication Date: 2021-05-18
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, triple-negative breast cancer lacks effective therapeutic targets available

Method used

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  • Polypeptide derived from mammary gland adipose tissue and anti-tumor application thereof
  • Polypeptide derived from mammary gland adipose tissue and anti-tumor application thereof
  • Polypeptide derived from mammary gland adipose tissue and anti-tumor application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Polypeptide PDBAG1-unmodified and PDBAG1 synthesis and analysis of biological parameters

[0024] The sequence of the polypeptide PDBAG1-unmodified is Ala Ser Lys Lys Val Cys Ile Val Gly SerGly Asn Trp Gly Ser Ala Ile Ala (SEQ ID NO.1), which is a polypeptide molecule composed of 18 amino acids, which naturally has acetylation at the N-terminus grooming. The main biological parameters of the peptide obtained through the online peptide analysis tool are as follows: the isoelectric point (Theoretical pI) is 9.32, and the molecular weight (Molecular weight, Mw) is 1748.03. Entrust Shanghai Keyeptide Biotechnology Co., Ltd. to synthesize the polypeptide with the sequence shown above by solid-phase method.

[0025] In order to optimize the performance of the polypeptide molecule and make it better enter the cell to function, we added a membrane-penetrating peptide fragment at its N-terminus: GRKKRRQRRRPPQQ, and modified the polypeptide sequence to: Gly Arg Lys Lys...

Embodiment 2

[0027] Example 2 Effects of Polypeptide PDBAG1 on the Viability of Triple Negative Breast Cancer Cells

[0028] The triple-negative breast cancer cell MDA-MB-231 was subjected to standard conditions (37°C, 5% CO 2 )nourish. On the first day, inoculate in a 96-well culture plate at a density of 2000 cells / 100ul medium / well; on the second day, treat with polypeptide PDBAG1-unmodified and PDBAG1 (set 10, 50μM high and low concentrations), control The group was treated with ultrapure water; after 24h, 48h, and 72h of polypeptide treatment, the medium containing 10% CCK-8 reagent was prepared respectively, the medium was replaced, and the culture was continued for 1h under standard conditions, and the absorbance value at 450nm wavelength was detected by a microplate reader , the growth curves are depicted in terms of OD values.

[0029] The result is as figure 1 As shown in A and B, the polypeptide PDBAG1-unmodified at a concentration of 50 μM can significantly inhibit the viabi...

Embodiment 3

[0030] Example 3 Distribution of FITC-labeled polypeptide PDBAG1 in cells

[0031] The triple-negative breast cancer cell MDA-MB-231 was subjected to standard conditions (37°C, 5% CO 2 )nourish. On the first day, they were seeded in 6-well culture plates at a density of 200,000 cells / 2ml medium / well; on the second day, the experimental group was treated with FITC fluorescently labeled PDBAG1 at a concentration of 30 μM, and the control group was treated with ultrapure water; After the polypeptide was treated for 24 hours, the cell entry of the polypeptide was observed using a fluorescence microscope.

[0032] The result is as figure 2 As shown, the polypeptide PDBAG1 can enter into triple-negative breast cancer cells, and is distributed in both the nucleus and cytoplasm, suggesting the possible mechanism of its function.

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Abstract

The invention discloses a polypeptide derived from mammary gland adipose tissue and anti-tumor application thereof. The amino acid sequence of the polypeptide PDBAG1 derived from the mammary gland adipose tissue is shown in SEQ ID No. 2. When the concentration of polypeptide molecules is low, the activity of breast cancer cells can be inhibited, apoptosis of the breast cancer cells can be promoted, and the anti-cancer effect is achieved mainly by damaging the glycolysis storage capacity and up-regulating active oxygen in tumor cells. When the dosage of the polypeptide PDBAG1 derived from the mammary gland adipose tissue is low, the remarkable cancer inhibition effect can be achieved, and the polypeptide is a small-molecule polypeptide, is easy to synthesize, efficient and low in toxicity and has a good anti-tumor effect.

Description

technical field [0001] The invention belongs to the field of anti-tumor, and relates to endogenous polypeptide of mammary gland fat tissue and its anti-tumor application. Background technique [0002] Breast cancer is a common malignant tumor in women. With the gradual enrichment of treatment methods, the survival prognosis of patients has been significantly improved, and the 5-year survival can reach 90%. However, triple-negative breast cancer is negative for estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2, and there is a lack of effective treatment methods in clinical treatment. Chemotherapy has always been the main method, which is still a difficult point in the treatment of breast cancer. and dilemma. Therefore, how to solve the problem of clinical treatment of triple-negative breast cancer has always been of great significance to breast cancer. [0003] Years of research have shown that triple-negative breast cancer can only be t...

Claims

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Application Information

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IPC IPC(8): C12N9/04C12N15/53C07K19/00A61K38/44A61K47/64A61P35/00A61P15/14
CPCA61K38/00A61P15/14A61P35/00C07K2319/10C12N9/0006C12Y101/01008
Inventor 吕明明陆迅李幸幸陆澄张敏陈飞
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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