The Investigation Of TIFA Expression In Lung Adenocarcinoma And The Mechanism In Promoting Malignant Biological Behavior

Foreword:Lung cancer is one of the leading cause of morbidity and mortality in the world.In lung cancer,the proportion of adenocarcinoma increases year by year,has become the main pathological type.But adenocarcinoma's quickly development,easy to metastasis and recurrence and the insensitivity to the radiation and chemotherapy lead to its poor prognosis is poorer.Although in recent years some new targeted therapies for the treatment of lung adenocarcinoma has improved,but the insensitivity or resistant still appear in the application.So exploring the lung adenocarcinoma new targets effectively,proving carcinogenic mechanism,play an important role for the treatment of lung adenocarcinoma.More and more experiences show that there are relationship between inflammation and cancer.Tumor necrosis factor receptor associated factor-interacting protein with a Forkhead-associated domain,abbreviated TIFA,is a kind of protein containing FHA structure domain,can interact with tumor necrosis factor receptor associated factor.It can participate in the downstream of the multiple tumor related signaling pathway.The TRAFs have been proved that play roles in some kinds of cancer,and also the protein that contain the FHA domain that do something with the malignant tumor.So we suggest that the TIFA may play some roles in cancer.But in the lung cancer,there has been no experiment about TIFA.Objective: To explore the TIFA expression in lung adenocarcinoma and explore its clinical significance.In the cell line research,observing the role of TIFA on proliferation,apoptosis,cell cycle and the invasion of lung adenocarcinoma.Through detecting the protein's exrpression in the downstream pathway after the changing of TIFA to invstigante the mechanisms that TIFA promotes cancer in the lung adenocarcinoma.Methods: 1.Collecting 116 cases of China medical university first affiliated hospital underwent surgical treatment of lung adenocarcinoma with complete follow-up data,51 cases of men,65 cases of women.By immunohistochemical method to detect TIFA gene expression of lung adenocarcinoma tissue adjacent to carcinoma tissues in the patients.Using the Chi-square test,single factor analysis and factor analysis to explore more TIFA the correlation between its expression and the prognosis of lung adenocarcinoma.Using Kaplan Meier method compare TIFA's different expression in impact on patients' survival.2.In the cell line research,using Western blot and Real-time PCR methods to detect the protein and m RNA's expression of TIFA in different cell lines.Screening TIFA high expression of lung adenocarcinoma cancer cell line.In lung adenocarcinoma H1975 and HCC827 cell line,TIFA silenced by the Lentivirus RNA interference.Through CCK8 method,flow cytometry instrument to detect the cell cycle and cell apoptosis,invasion and scratch test,verify the TIFA's role in biological behavior of lung adenocarcinoma cancer cell.3.In lung adenocarcinoma H1975 and HCC827 cell line,through the Lentivirus mediated transfection to make TIFA overexpression of silence.Using Western blot to detection nf-kappa B signaling pathway activation,and the changes of some protein for example the p53 and Bcl-2,etc.Result: 1.In the clinical patients' research,immunohistochemical staining confirmed that in 116 cases of lung adenocarcinoma and adjacent tissue.In cancerous tissue the TIFA positive expression number is 63(54.3%),but in the tissue adjacent to carcinoma,the positive expression rate is only 30.1%,and the difference is statistically significant.The expression of TIFA is related to the degree of differentiation,TNM staging and survival of the lung adenocarcinoma.Multiple factors analysis showed that positive expression of TIFA is independent risk factors for the patients' survival of lung adenocarcinoma.Kaplan-Meier survival analysis showed that TIFA positive expression patients obviously have shorten lifetime.2 In lung adenocarcinoma cells,the expression of TIFA is generally higher than normal cells.In lung adenocarcinoma H1975 and HCC827 cell line,using lentivirus mediated TIFA silence,compared with the control group,the cell proliferation was obviously inhibited in the silence group(CCK8 method).Using flow cytometry analysis to detect the cell cycle,in TIFA silence group,the G0 / G1 phase ratio was increased in two cell lines,suggesting that silencing TIFA can have the ability to make cell cycle arrest.The apoptosis ratio of TIFA silent group also increased significantly,indicating that TIFA has the ability to regulate the apoptosis of cell.After the silence of TIFA,H1975 and HCC827 cell invasion and migration ability decreased compare to the control group,suggesting that the TIFA can regulate the invasion and migration ability of lung adenocarcinoma cell line.3.In lung adenocarcinoma H1975 and HCC827 cell line,using the lentivirus tranfection to mediate TIFA overexpression or silence.Founding that in lung adenocarcinoma cells,TIFA can activate the nf-kappa B signaling pathway.At the same time,TIFA expression changes also changed the expression of downstream protein for example the p53,Bcl-2,p21,Cyclin D1,etc.Indicating that TIFA is associated with the malignant biological behavior of lung adenocarcinoma.Conclusion: 1.TIFA expresses positively in lung adenocarcinoma tissue,its expression is related to the degree of tumor differentiation,TNM staging and the survival of lung adenocarcinoma.2.TIFA's expression is maybe the independent risk factors for survival in patients with lung adenocarcinoma.3.In lung adenocarcinoma cells,the expression of TIFA is generally higher.4.TIFA can promote the cell proliferation,invasion and migration of lung adenocarcinoma,TIFA's expression can promote cell cycle progression and apoptosis inhibition 5.TIFA in lung adenocarcinoma can activate the nf-kappa B pathway and affect the downstream protein expression to promote malignant biological behavior of tumors.