The Mechanism Of Corosolic Acid On Proliferation,Invasion And Chemotherapy Resistance Regulating In Non-small Cell Lung Cancer

Purpose: To elucidate the regulatory effects of korosolic acid on proliferation,invasion and chemotherapy resistance of NSCLC,and to reveal its regulatory mechanism.Methods: PC9 and A549 cell lines were employ,CCK8,EDU,plate cloning and other techniques were used to detect the change of proliferation ability of NSCLC cells with or without colosolic acid treatment.Wound-healing and transwell migration analysis were used to detection the effects of colosolic acid on cell invasion and migration.Cell apoptosis and cell cycle were detected by flow cytometry using apoptosis and cycle kit with or without colosolic acid detection.Cisplatin resistant strain A549-DDP was used to study the effect of corosolic acid on chemotherapy resistance of cells.Mitochondrial oxidative stress kit,lipid peroxidation detection kit and mitochondrial membrane potential detection kit were used to detect the effects of korosolic acid on oxidative stress and mitochondrial damage.BALB/C nude mice were used to construct tumor-bearing mouse model and caudal vein metastasis model to detect the effects of corosolic acid on tumor growth,metastasis and chemotherapy resistance.Metabonomics and high-throughput sequencing were used to detect NSCLC cell metabolism and related gene expression before and after treatment with colosolic acid.Results: The results of in vitro and in vivo experiments showed that compared with the control group,the proliferation,migration and invasion of NSCLC cells were significantly inhibited by colosolic acid treatment.The IC50 of cell proliferation and migration related to A549 and PC9 were 27.5 μg/m L and 15 μg/m L,respectively,and the IC50 of cell proliferation and migration related to PC9 were 12.5 μg/m L and 7.5μg/m L,respectively.Colosolic acid treatment caused cell division to stagnate in the G2/M phase.Colosolic acid promoted apoptosis and increased the chemosensitivity of NSCLC cells to cisplatin.Immunofluorescence assays showed that korosolic acid treatment increased mitochondrial and liposome oxidative stress(ROS)and damaged mitochondrial membrane potential in NSCLC cells.Metabonomics analysis showed that korosolic acid treatment inhibited the expression of reduced glutathione.Ferrostatin 1,an ferroptosis inhibitor,reversible conversion to corosolic acid induced apoptosis;High-throughput sequencing and RT-q PCR detection showed that corosolic acid inhibited the expression of GPX2,TPX2,CCNB1,CDK1,and PI3K/Akt signal,but promoted the expression of pro-apoptotic signaling protein Caspase-3.Conclusions: The study confirmed that Russell acid inhibits cell proliferation,invasion and chemotherapy drug resistance,and further studies confirmed that secco barroso acid damaged GPX2 mediated GSH REDOX system stability,liposome increased oxidative stress and mitochondrial oxidative stress,and at the same time division barroso acid treatment also reduces the TPX2 expression,thus inhibiting the PI3K/Akt signal,leading to cell apoptosis.The accumulation of ROS can dissociate the CCNB1/CDK1 complex,induce G2/M cell cycle arrest,and eventually lead to cell apoptosis.In summary,our study found that natural triterpene kroxoic acid can reduce the proliferation,invasion and chemotherapy resistance of NSCLC by activating mitochondrial and liposome oxidative stress.