A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells

Nereis active protease (NAP) is a novel fibrinolytic active serine protease from the polychaete Nereis virens. In this study, NAP was purified from Nereis virens and the effects of NAP on human lung cancer cells were investigated. Our results indicated that NAP inhibited the proliferation and induce...

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Main Authors: Yunping Tang, Fangmiao Yu, Guomei Zhang, Zuisu Yang, Fangfang Huang, Guofang Ding
Format: Article
Language:English
Published: MDPI AG 2017-07-01
Series:Molecules
Subjects:
Online Access:https://www.mdpi.com/1420-3049/22/7/1123
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spelling doaj-e47cf527937d453cbe007f27f282b4762020-11-25T00:57:51ZengMDPI AGMolecules1420-30492017-07-01227112310.3390/molecules22071123molecules22071123A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer CellsYunping Tang0Fangmiao Yu1Guomei Zhang2Zuisu Yang3Fangfang Huang4Guofang Ding5School of Food and Pharmacy, Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, Zhejiang Ocean University, 1st Haidanan Road, Changzhi Island, Lincheng, Zhoushan 316022, ChinaSchool of Food and Pharmacy, Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, Zhejiang Ocean University, 1st Haidanan Road, Changzhi Island, Lincheng, Zhoushan 316022, ChinaSchool of Food and Pharmacy, Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, Zhejiang Ocean University, 1st Haidanan Road, Changzhi Island, Lincheng, Zhoushan 316022, ChinaSchool of Food and Pharmacy, Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, Zhejiang Ocean University, 1st Haidanan Road, Changzhi Island, Lincheng, Zhoushan 316022, ChinaSchool of Food and Pharmacy, Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, Zhejiang Ocean University, 1st Haidanan Road, Changzhi Island, Lincheng, Zhoushan 316022, ChinaSchool of Food and Pharmacy, Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, Zhejiang Ocean University, 1st Haidanan Road, Changzhi Island, Lincheng, Zhoushan 316022, ChinaNereis active protease (NAP) is a novel fibrinolytic active serine protease from the polychaete Nereis virens. In this study, NAP was purified from Nereis virens and the effects of NAP on human lung cancer cells were investigated. Our results indicated that NAP inhibited the proliferation and induced apoptosis of H1299 cells in a time- and dose-dependent manner. The loss of mitochondrial membrane potential, the activation of Bax and cleaved-caspase 3/9, the release of cytochrome C, and the suppression of Bcl-2 and poly-ADP ribose polymerase were observed in NAP-treated H1299 cells by flow cytometry and Western blotting. Moreover, the expression levels of Bax and Bcl-2 mRNA were determined by real-time quantitative polymerase chain reaction and the Bax/Bcl-2 expression ratio was increased in the NAP-treated cell lines. The results indicated that NAP-induced apoptosis may be related to mitochondria mediated apoptosis and occurs through caspase-dependent pathways. Then, the effects of NAP on tumor growth in animal models were observed, where 5 or 10 mg/kg of NAP noticeably reduced tumor volume and weight and increased apoptosis as determined by Western blotting when compared to the negative control group. Therefore, our findings suggest that NAP could be a hopeful anticancer medicine for its propensity to inhibit growth and induce of apoptosis in human lung cancer cells.https://www.mdpi.com/1420-3049/22/7/1123Nereis virensserine proteaselung cancerapoptosismitochondrial dysfunction
collection DOAJ
language English
format Article
sources DOAJ
author Yunping Tang
Fangmiao Yu
Guomei Zhang
Zuisu Yang
Fangfang Huang
Guofang Ding
spellingShingle Yunping Tang
Fangmiao Yu
Guomei Zhang
Zuisu Yang
Fangfang Huang
Guofang Ding
A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells
Molecules
Nereis virens
serine protease
lung cancer
apoptosis
mitochondrial dysfunction
author_facet Yunping Tang
Fangmiao Yu
Guomei Zhang
Zuisu Yang
Fangfang Huang
Guofang Ding
author_sort Yunping Tang
title A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells
title_short A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells
title_full A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells
title_fullStr A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells
title_full_unstemmed A Purified Serine Protease from Nereis virens and Its Impaction of Apoptosis on Human Lung Cancer Cells
title_sort purified serine protease from nereis virens and its impaction of apoptosis on human lung cancer cells
publisher MDPI AG
series Molecules
issn 1420-3049
publishDate 2017-07-01
description Nereis active protease (NAP) is a novel fibrinolytic active serine protease from the polychaete Nereis virens. In this study, NAP was purified from Nereis virens and the effects of NAP on human lung cancer cells were investigated. Our results indicated that NAP inhibited the proliferation and induced apoptosis of H1299 cells in a time- and dose-dependent manner. The loss of mitochondrial membrane potential, the activation of Bax and cleaved-caspase 3/9, the release of cytochrome C, and the suppression of Bcl-2 and poly-ADP ribose polymerase were observed in NAP-treated H1299 cells by flow cytometry and Western blotting. Moreover, the expression levels of Bax and Bcl-2 mRNA were determined by real-time quantitative polymerase chain reaction and the Bax/Bcl-2 expression ratio was increased in the NAP-treated cell lines. The results indicated that NAP-induced apoptosis may be related to mitochondria mediated apoptosis and occurs through caspase-dependent pathways. Then, the effects of NAP on tumor growth in animal models were observed, where 5 or 10 mg/kg of NAP noticeably reduced tumor volume and weight and increased apoptosis as determined by Western blotting when compared to the negative control group. Therefore, our findings suggest that NAP could be a hopeful anticancer medicine for its propensity to inhibit growth and induce of apoptosis in human lung cancer cells.
topic Nereis virens
serine protease
lung cancer
apoptosis
mitochondrial dysfunction
url https://www.mdpi.com/1420-3049/22/7/1123
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