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Polypeptide Inducing Prostate Cancer Cell Apoptosis and Its Application

A cancer cell, prostate technology, applied in the field of medicine and biology, can solve the problems of reduced autophagy, toxicity, and enhanced anti-apoptotic ability of tumor cells, and achieves good therapeutic effect.

Active Publication Date: 2020-08-21
中美赛尔生物科技(广东)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Based on a domain of PSMA that can recognize drugs outside the membrane, some researchers designed and synthesized a series of methotrexate conjugates that can be activated by PSMA, prompting PSMA to play the role of exopeptidase and release methotrexate , to play a tumor-killing effect, but the expected effect may not be achieved due to reasons such as too large a conjugate molecule and the toxicity of methotrexate
[0008] On the other hand, the deletion of the autophagy-related gene Beclin1 is the mechanism for the reduction of autophagy and the enhancement of anti-apoptosis ability of tumor cells

Method used

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  • Polypeptide Inducing Prostate Cancer Cell Apoptosis and Its Application
  • Polypeptide Inducing Prostate Cancer Cell Apoptosis and Its Application
  • Polypeptide Inducing Prostate Cancer Cell Apoptosis and Its Application

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Experimental program
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Effect test

Embodiment 3

[0039] Take LNcap cells and adjust the cell concentration to 1x10 6 pcs / ml. Inoculate 100 μl at a concentration of 1x10 in Cornng Transwell plate chambers 6 individual / ml LNcap cells, and added different mass concentrations (0, 2, 10, 50, 100 μg / mL) of polypeptides respectively. Add 500 μl of RPMI1640 medium containing 40% FBS to the lower chamber, place the plate in the incubator for 18 hours, remove the lower chamber and discard the medium in the upper chamber, fix it with 4% paraformaldehyde solution, stain with crystal violet and put it under an inverted microscope. Cells moved to the lower layer of the microporous membrane were counted under the 200X magnification finder frame. An average of 10 fields of view were counted per chamber. The migratory capacity of the cells was analyzed by averaging.

[0040] like figure 1 It is shown that the polypeptide of the present invention can effectively inhibit the migration ability of LNcap cells in a dose-dependent manner. Wit...

Embodiment 4

[0042] LNcap cells at 5 x 10 5 The cells / well were inoculated into 6-well plates, cultured at 37°C and 5% CO2 for 24 hours, and then added with different mass concentrations (0, 2, 10, 50, 100 μg / mL) of polypeptides. After culturing for 48 hours, the cells were collected, RNA was extracted, and reverse transcribed into cDNA. Using GAPDH as an internal reference, the expression of apoptosis inhibitory gene Bcl-2 and autophagy-related gene Atg5 was detected by qPCR.

[0043] The primer sequences are:

[0044] GAPDH Forward: 5'-GGAGCGAGATCCCTCCAAAAT-3',

[0045]GAPDH Reverse: GGCTGTTGTCATACTTCTCATGG-3';

[0046] Bcl-2 Forward:GGTGGGGTCATGTGTGTGG-3’,

[0047] Bcl-2 Reverse:CGGTTCAGGTACTCAGTCATCC-3';

[0048] Atg5 Forward: 5'-AAAGATGTGCTTCGAGATGTGT-3',

[0049] Atg5 Reverse: 5'-CACTTTGTCAGTTACCAACGTCA-3'.

[0050] The result is as figure 2 As shown, the polypeptide of the present invention can accelerate the apoptosis of LNcap cells by down-regulating the expression of the ...

Embodiment 5

[0052] PSMA can activate the PI3K / AKT signaling pathway by up-regulating the phosphorylation level of AKT, and increase the proliferation and metastasis of prostate cancer cells.

[0053] The prostate cancer cell line LNcap cells were plated at 5 × 10 5 The density of cells / well was inoculated into 6-well plates, and after culturing for 24 hours at 37°C and 5% CO2, polypeptides of different mass concentrations (0, 2, 10, 50, 100 μg / mL) were added, and the cells were collected after culturing for 48 hours.

[0054] After fully washing with 4°C pre-cooled PBS three times, 200 μL of RIPA lysis buffer containing protease inhibitors and phosphatase inhibitors at 4°C was added to resuspend the cells and lysed on ice for 20 min. The supernatant protein solution was collected by centrifugation, and 150 μl of the protein solution was added to 37.5 μl of 5× protein buffer, boiled at 100°C for 5 min, and stored at 4°C for later use.

[0055] The prepared protein solution was added to th...

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Abstract

The invention relates to a polypeptide for inducing apoptosis of prostate cancer cells and application thereof in preparation of drugs for treating prostate cancer of subjects.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a polypeptide that induces apoptosis of prostate cancer cells and its use in the preparation of a medicament for treating prostate cancer in a subject. Background technique [0002] Prostate carcinoma is an epithelial malignant tumor that occurs in the prostate, with high morbidity and mortality, and is one of the most common malignant tumors in men. The etiology of prostate cancer is still unclear, and may be related to various factors such as heredity and environment. Like other human tumors, inflammation and immunosuppression play an important role in the occurrence and development of prostate cancer. [0003] Current studies suggest that host components that interact with tumors may be involved in the formation of an immunosuppressive microenvironment for the occurrence and development of prostate cancer. A large number of evidences show that prostate cancer is immune-re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00A61K38/16A61P35/00
CPCA61K38/00C07K14/00
Inventor 李陶王旭林词雄林洁璇朱刚
Owner 中美赛尔生物科技(广东)有限公司
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