Study On The Mechanism Of Anti-lung Cancer Of Nereis Virens Protease (NAP) Based On PI3K/AKT/mTOR And ERK/MAPK Signaling Pathways

Lung cancer,as one of the most common malignant tumor diseases in the world,is a serious threat to human health and life due to its extremely high morbidity and mortality.As one of the two subtypes of non-small cell lung cancer,most is not confirmed by surgical treatment of late stage or transfer stage,only by means of radiotherapy or chemotherapy treatment,such as cisplatin,carboplatin,and etoposide chemotherapy drugs,but the drug toxicity is larger,and the long-term use of easy to produce drug resistance,so looking for a kind of high efficient and low toxicity anti-cancer drugs urgently demand.Because sea creatures living in the unique high salt,high pressure,low temperature,dark,hypoxia and strong corrosive environment,the amino acid composition of enzymes or sequence and terrestrial protein is different,in a wide variety of Marine protease in the sequence of amino acids,with many potential bioactive amino acid sequence of the activity of protease.Therefore,based on the PI3K/AKT/mTOR and ERK/MAPK signaling pathways,this master’s thesis is to investigate the mechanism of napa protease(NAP)in the fight against lung cancer.Firstly,transcriptome sequencing method was used for the study.According to the research results,MAPK and PI3K/AKT/mTOR signaling pathways were selected as the research direction of this paper.Then,through cell scratch,cell cycle and Western Blotting experiments in vitro,it was found that NAP may induce apoptosis by inhibiting the PI3K/AKT/mTOR pathway.We then detected changes in p-erk,p-akt and cleaved PARP proteins at different time periods,and found that key protein expression in the NAP inhibition pathway occurred before apoptosis.At the same time,PI3K/AKT/mTOR and ERK/MAPK signaling inhibitors LY294002 and PD98059 were used to act on the cells,and it was found that the former enhanced the phosphorylation of NAP induced ERK and AKT,while the latter had no significant effect on the phosphorylation of NAP induced ERK and AKT.We speculated that PI3K/AKT/mTOR was one of the potential signaling pathway mechanisms for NAP induced apoptosis of H1299 cells.Finally,in vivo experiments were conducted to study the effect of NAP on transplanted tumors in nudemice.It was found that NAP showed significant inhibition effect on transplanted tumors in nude mice.Compared with the cisplatin group,the inhibition effect of NAP group was low but the toxic and side effects were not significant.Therefore,NAP showed a significant inhibitory effect on the PI3K/AKT/mTOR pathway in H1299 tumor-bearing nude mice transplanted tumor,inhibited the activation of the PI3K/AKT/mTOR signaling pathway,affected cell proliferation,and thus inhibited tumor growth.