Mechanism Of C-Raf Kinase Binding Protein SMC3 And PP2A A? In Lung Carcinogenesis

Cancer is a malignant tumor originating in epithelial tissue,and lung cancer is the most common type of malignancy,the world's highest mortality rate and the highest incidence of cancer,it gives people life and the body a great threat.Therefore,the study of the pathogenesis of lung cancer,lung cancer to find a new target is even more urgent.Raf kinase(Rapidly Accelerated Fibrosarcoma),including A-Raf,B-Raf and C-Raf three subtypes,Raf kinase mutation is one of the main causes of cell cancer,A-Raf mutations can lead to ovarian and colorectal cancer,B-Raf mutations and neurological carcinogenesis,skin cancer and thymoma-associated,C-Raf mutations are associated with small cell lung cancer,and the expression of C-Raf protein in lung cancer tissues Significantly increased,therefore,regulation of C-Raf kinase and its binding protein has become an important means of treatment of lung cancer.However,most cancer treatments focus on a single target or a single mutation,lacking ahigh degree of understanding of the dynamic cellular signaling network from the height of thesystem and an in-depth understanding of its intrinsic mechanisms.In this study,we investigated the role of key nodes in the tumor cell signaling network in tumorigenesis with the C-Raf central protein interaction network.We used protein cross-group and high-throughput mass spectrometry to systematically identify C-Raf interacting proteins SMC3(Structural maintenance of chromosomes protein 3)and PP2A A?(protein phyosphatase2A Aa).SMC3 is an important component of adhesin,SMC3 and SMC1A subunit through its hinge domain dimerization to form 'V' shape,this 'V' and the bottom of the RAD21 subunit connected to form a closed trilateral ring structure,this The circular structure can encircle the sister chromosome to mediate the adhesion of sister chromosome connexin and play a key role in gene regulation.SMC3 can be acetylated in S phase to establish the adhesion of chromatin-loaded fibronectin,which affects Cell cycle.Protein phosphatase PP2A Aa is one of the important components of protein phosphatase PP2A,PP2A is a serine threonine protein phosphatase,which is very high in most tissues and cells,It is involved in the replication of DNA,gene transcription,cell differentiation and apoptosis,oncogene transformation and a series of cell biology activities,in addition it also has a certain relationship with the occurrence and development of tumor,Mutation of PP2A Aa protein is closely related to the occurrence of lung cancer and uterine cancer.We constructed the high expression plasmids of SMC3 and PP2A Aa protein(pcDNA3.1-SMC3/PP2A,Aa),knockdown plasmid(pSUPER-SMC3/PP2A,Aa)and knockout plasmid(px459-SMC3/PP2A,A),Stably transforming into non-small cell lung cancer A549 cells,we successfully constructed a high expression and knockdown stable cell line of SMC3/PP2A Aa.Finally,we detected the proliferation of A549 cells by CCK8 method and found that the proliferation ability of A549 cells stabilized with pSUPER-SMC3 was significantly lower than that of control group and negative test group(pSUPER)(P<0.01),suggesting that the stabilization of pSUPER-SMC3 inhibited the proliferation of A549 cells;The migration of A549 cells after pSUPER-SMC3 was detected by scratches and Transwell chamber test,The number of cells in the experimental group(pSUPER-SMC3)was significantly lower than that in the blank control group and the negative control group(P<0.05),which indicated that down-regulation of SMC3 protein expression could inhibit the migration of A549 cells;Cell viability and stemness were measured by cell pellet assay,Found that the experimental group cells into the ball number(19.66+3.055)were significantly less than the control group(54.333+4.041)and control group(54.667+3.786)(P<0.05),and the blank control group and negative control group cells count was not significant(P>0.05),prove that the expression of SMC3 protein of A549 cells into the ball to reduce the number of sternness,weakened.In general,we selected and identified C-Raf binding protein SMC3 and PP2A Aa,and successfully constructed SMC3 and PP2A Aa high expression,knockdown and knockout plasmid,and then transferred into A549 cells,successfully constructed SMC3 and PP2A Aa overexpressing cell lines and knockdown cell lines.Second,it is proved that the silencing of SMC3 will inhibit the migration and ball ability of A549 cells and reduce the dryness of cells.These are important biological functions of cancer cell survival and tumor development.Therefore,SMC3 is expected to become a new biological Signs and treatment of drug targets.