Study On The Mechanism Of C-Raf Kinase Binding Protein KAP1 In Lung Cancer

Lung cancer is the leading cause of all cancer-related deaths.In fact,the death of individual lung cancer exceeds the sum of the three most common cancer deaths(colon,breast and prostate).The poor prognosis of lung cancer is due to the lack of early diagnosis of any significant symptoms during the progression of early disease.In addition,a variety of genetic changes identified in NSCLC,including mutations,deletions,deletions and fusions,exacerbate abnormalities in signaling pathways and physiological activities.Therefore,to explore an effective target for urgent treatment of lung cancer.Several decades of studies have shown that the mitogen-activated protein kinase(MAPK)pathway transduces many different signals,resulting in a wide range of cellular responses,including cell proliferation,differentiation,survival,migration,neuronal function and immune response.Raf-MEK-ERK pathway is a trunk pathway of cell signaling,which is a retroviral oncogene that regulates cell differentiation,transformation,growth and apoptosis,abnormal regulation is one of the main cell cancer.The physiological regulation of RAF kinases is complex and involves several steps,including the interaction between protein and protein,phosphorylation,dephosphorylation and conformational changes.And C-RAF as the main hub protein in the MAPK pathway,its function and binding protein play an important role in the development and progression of lung cancer.KAP1 is a member of the human TRIM gene family and is highly correlated with the other three TRIM proteins(TIF1?,TIF1? and TIF1?).KAP1 is a key regulator of normal development and differentiation;mice lacking KAP1 die before pretreatment and mice with specific absence of KAP1 in adult forebrain show a higher level of anxiety and stress-induced learning and memory It changes.KAP1 is also involved in maintaining pluripotency,which is necessary for terminal differentiation of mouse embryonic stem cells and has been associated with the promotion and inhibition of differentiation of different adult cell types.KAP1 play a key role in proliferation and differentiation in normal and tumor cells.In this study,we found that C-RAF could specifically bind to KAP1.The expression of KAP1 in lung cancer tissues and adjacent normal lung tissues was detected by experiments.It was found that KAP1 was highly expressed in lung cancer tissues.In cell experiments,CRAPP/cas9 was successfully used to successfully knock out KAPI in lung cancer A549 cell line and rescue assay.The sensitivity of cisplatin and 5-fluorouracil(5-FU)was significantly increased in KAP1 cells after knockout of KAP1,and it was found that low concentration of 5-FU cells did not promote the apoptosis of A549 cells,and the cells were knocked out after knockout of KAP1.The cells were detected by western blot.Resistant cells and knockout KAP1 cells were identified to be activated by knockout KAP1.Scaffold and transwell showed that the cell migration was significantly inhibited after knockout of KAP1.The results showed that the knockout rate of KAP1 was significantly decreased,and that of western blot showed that the cell viability was significantly decreased;the soft agar clone was found to knock out KAP1 The number of clones was significantly reduced;the formation of blood vessels found knocked down KAP1,the angiogenic ability decreased significantly;EMT transformation experiments found that knockout KAP1,the cell transformation ability was significantly reduced;loss of cell cycle showed significant inhibition of cell cycle;Indicating that the volume and weight of transplanted tumor in nude mice after knockout of KAP 1 were significantly decreased;the weight of nude mice increased significantly;the proliferation of transplanted tumor in AKO group Degree significantly slower than the A549 group.Compared with AKO group,the tumor cells were irregular,the nucleus hypertrophy was blue,and the mitotic phase was common in the A549 group.In this study,KAP1 plays an important role in lung cancer cell line A549.After cell knockout,cell proliferation,cell migration,invasion and migration ability,vascular cell formation and cell cycle are inhibited,and abnormal expression of ERK in A549 cells is improved.FU sensitivity.At present,the role of C-RAF and KAP 1 in the development of lung cancer is still very small,this study for their role in the pathogenesis of lung cancer were initially explored for the clinical treatment of lung cancer to provide theoretical basis and new targets.