Study On The Mechanism Of HSP90AA1 Promoting Invasion And Metastasis Of Lung Adenocarcinoma Through AKT Pathway

Objective:Lung adenocarcinoma is A malignant tumor with high morbidity and mortality.Heat shock protein 90 alpha family class A member 1 HSP90AA1 is highly expressed in lung adenocarcinoma metastasis tissues.However,the mechanism of its effect on lung adenocarcinoma metastasis is rarely reported.This study aims to study the mechanism of HSP90AA1 affecting the proliferation,migration and invasion of lung adenocarcinoma cells,and to explore the mechanism of HSP90AA1 gene promoting the invasion and metastasis of lung adenocarcinoma through the AKT(protein kinase B)pathway,so as to provide basis for the prevention and treatment of lung adenocarcinoma and the search for new biomarkers.Methods:1.The expression of HSP90AA1 in tumors was analyzed using The Cancer Genome Atlas TCGA public database;2.Immunohistochemistry was used to detect the expression of HSP90AA1 protein in lung adenocarcinoma tissues,normal lung tissues,primary lung adenocarcinoma tissues and metastatic lung adenocarcinoma tissues,and to analyze the relationship between HSP90AA1 protein expression and clinicopathological features;3.Lung adenocarcinoma cells were infected with HSP90AA1-sh RNA lentivirus vector,and lung adenocarcinoma cells PC-9 HSP90AA1 low expression cell model was constructed;Lung adenocarcinoma cells HCC827 HSP90AA1 overexpression cell model was constructed by overexpression lentivirus vector HSP90AA1,and lung adenocarcinoma cells were infected with HSP90AA1;The changes of proliferation,migration and invasion of lung adenocarcinoma cells after knockdown and overexpression of HSP90AA1 were verified by cell cloning,CCK-8 cell proliferation,cell scratch assay and Transwell cell migration and invasion;Realtime-PCR and Western Blot were used to detect the expression of HSP90AA1 m RNA and protein in each transfected cell to verify the transfection efficiency and screen out stable cell lines;Western Blot was used to detect the expression of HSP90AA1,AKT,p-AKT and epithelial-mesenchymal Transition(EMT)related proteins in AKT pathway and HSP90AA1;4.In order to evaluate the effect of HSP90AA1 on tumor growth,nude mice were transplanted with subcutaneous tumor of the back,and the subcutaneous tumor model of the nude mice was established;The volume and weight of the transplanted tumor were observed periodically.Results:1.The expression of HSP90AA1 in lung adenocarcinoma tissues was higher than that in normal lung tissues,while the expression in metastatic lung adenocarcinoma tissues were higher than that in primary lung adenocarcinoma tissues;2.Human lung adenocarcinoma cells PC-9 and HCC827 expressed HSP90 AA 1 in different degrees;3.HSP90AA1-sh RNA lentivirus down-regulated the expression level of HSP90AA1 in PC-9 cells,and overexpression of HSP90AA1 lentivirus vector up-regulated the expression level of HSP90AA1 in HCC827 cells;Knockdown of HSP90AA1 significantly inhibited the proliferation,migration and invasion abilities of PC-9 cells,while the proliferation,migration and invasion abilities of HSP90AA1 and HCC827 cells were significantly enhanced;4.Knockdown of HSP90AA1 in lung adenocarcinoma cells significantly reduced the expression level of p-AKT in AKT pathway,while the expression level of AKT did not change significantly;The expressions of EMT-related proteins snail,MMP9 and Vimentin also decreased to varying degrees,while the expression of E-cadherin was up-regulated;Overexpression of HSP90AA1 up-regulated the expression of p-AKT,snail,MMP9 and Vimentin,and decreased the expression of E-cadherin;5.Through subcutaneous tumor transplantation experiments in nude mice,we found that knocking down HSP90AA1 reduced the volume and weight of tumors,while overexpression of HSP90AA1 promoted the proliferation of tumor cells.Conclusion:1.High expression of HSP90AA1 in metastatic lung adenocarcinoma tissues;2.Down-regulating the expression of HSP90AA1 in PC-9 cells could inhibit the proliferation,migration and invasion of cells,while up-regulating the expression of HSP90AA1 in HCC827 cells could promote the proliferation,migration and invasion of cells;3.HSP90AA1 can affect the invasion and migration of lung adenocarcinoma cells and promote tumor metastasis by inducing EMT through AKT pathway.