The Mechanisms Of PRSS3 In Mediating Proliferation And Metastasis Of Lung Cancer

Section 1:The Role and Mechanism of PRSS3-SPINKs-KLK5 Pathway in Mediating the Proliferation and Metastasis of Lung AdenocarcinomaObjective:Based on clinical,celluar,and bioinformatic studies,we aimed to explore the role of PRSS3 in mediating proliferation and metastasis of lung adenocarcinoma.Methods:We searched several bioinformatic data base and analyzed the PRSS3 expression level in lung adenocarcinoma patients.We also explored the correlation of PRSS3 expression with survival of lung adenocarcinoma patients.We transfected PC9,HCC827 and A549 cells with PRSS3 knock down lentivirus,and investigated the proliferation and invasion abilities of cells compared with the non-target control groups.We detected SPINK5/6,KLK5 substrates and KLK5 levels by western blot,and also detected the levels of KLK5 substrates after PRSS3 knock down.We constructed mouse othotopic model with PRSS3 knock down PC9 cells and studied the effects of PRSS3 knock down in vivo.We also looked into the effects of mesotrypsin inhibitors in mediating proliferation and metastasis of PC9 cells.Results:We found that PRSS3 expression was elevated in adenocarcinoma patients by analyzing the data on Oncomine,and K-M plotter online analysis showed that PRSS3 high expression was significantly associated with worse survival.Our data also showed that after PRSS3 knock down,the proliferation and/or invasion abilities of A549,HCC827 and PC9 cells were inhibited.After PRSS3 knock down,the KLK5 substrates VTN,IGFBP3 and collagen1 were elevated,while the SPINK5/6 levels were elevated,as judged by western blot.Co-IP showed that SPINK5/6 could directly react with KLK5.We also found that the KLK5 activity was reduced after PRSS3 knock down.We found that PRSS3 knock down inhibited tumor genesis and proliferation in the above mouse model.Mesotrypsin inhibitor could significantly inhibit the proliferation and metastasis of PC9 cells.After KLK5 and PRSS3 knock down,the p-AKT and SNAIL1 levels were reduced,which indicated that the proliferation and EMT levels were reduced in H520 cells.Conclusion:Based on the above clinical,celluar,and bioinformatic analysises,we found that PRSS3 could promote proliferation and metastasis of lung adenocarcinoma,so as to affect the prognosis of patients.The PRSS3-SPINK5-KLK5 pathway can mediate the proliferation and metastasis of lung adenocarcinoma.PRSS3 could be a new treatment therapeutic target for lung adenocarcinoma.Section 2:Expoloration of PRSS3's Role in Mediating Proliferation and Metastasis of Lung Squamous Cells CarcinomaObjective:We found that PRSS3 promoted proliferation and metastasis of lung adenocarcinoma through PRSS3-SPINKs-KLK5 pathway to affect cell phenotypes.In this section we aimed to investigate the function of PRSS3 in lung squamous cell carcinoma.Methods:We searched the Oncomine database and analyzed the PRSS3 expression level in lung squamous cell carcinoma patients.We searched the K-M plotter online database and explored the correlation of PRSS3 expression with survival of lung squamous cell carcinoma patients.We detected the SNAIL1 and p-AKT/AKT levels in cell lysates of H520 cells transfected with PRSS3 knock down lentivirus,and compared with the non-target control groups to explore the downstream pathway assiociated with proliferation and metastasis change.Results:Our data also showed that after PRSS3 knock down,the proliferation and metastasis of PRSS3 was inhibited in H520 cells.Similar phenomenon was seen in KLK5 knock down H520 cells.The SNAIL 1 and p-AKT level was reduced after KLK5 and PRSS3 knock down,which indicated that the proliferation,EMT level and cell metastasis ability was reduced after PRSS3 knock down.Conclusion:We showed that after PRSS3 knock down in H520 cells,the cell proliferation and metastasis was inhibited.PRSS3 could possibly mediate the H520 proliferation and metastasis by regulating KLK5 activity,and detailed machenisms of PRSS3 in mediating H520 proliferation and metastasis need further investigation.