Abstract
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.
Highlights
Lung cancer is one of the most common and fatal cancers in the world [1,2]
Nereis active protease (NAP) was purified from Nereis virens through ammonium sulfate precipitation, anion exchange
NAP was purified from Nereis virens through ammonium sulfate precipitation, anion exchange chromatography, and gel chromatography
Summary
Recent studies show that almost one third of all cancer-related death in males and one fourth of all cancer death in females are due to the cause of lung cancer. The incidence and mortality of lung cancer in China has increased because of smoking and increased air pollution [5,6]. Surgery, radiation, chemotherapy and targeted therapy are used in lung cancer treatment [7,8,9,10]. Resistance to anticancer drugs has been observed and normal cells may be destroyed by the side effects of anticancer drugs [11,12]. It is necessary to study and develop more effective and low-toxicity natural anticancer drugs
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