| BackgroundThere are many types of malignant tumors in the world,but lung cancer is common to us.Lung cancer is the highest among the types of cancer that causes death,both in men and in women.Non small cell lung cancer is in a large proportion in all types of lung cancer,and about 4/5 of all types of lung cancer are non small cell lung cancer.Some scholars believe that cells can not control growth in the lung tissue,and may lead to the occurrence of metastasis.Like other cancers,the occurrence of lung cancer is also the two type of gene abnormality,that is,tumor suppressor genes and oncogenes.The function of oncogenes or tumor suppressor genes is out of order,which leads to the occurrence of lung cancer.Among the factors that induce lung cancer,smoking is an important cause.In recent years,despite the continuous progress and improve the level of diagnosis and treatment of medical technology,but due to the lack of early symptoms and lung lesions often accumulate in the deepest parts of the lungs.When they are diagnosed,they are mostly in the middle and late stages,or have metastasized,and patients missed the best treatment period,the 5-year survival rate of lung cancer patients has not been significantly improved,which has caused a serious impact on human health.Programmed cell death factor 4,also known as programmed cell apoptosis factor 4,is a well known tumor suppressor gene,which can be expressed in nucleus and cytoplasm.It plays an important role in transcription,translation and apoptosis.It has been reported in a variety of tumors: it is in a state of low expression,such as in lung cancer,colon cancer,breast cancer,esophageal cancer,stomach cancer,and ovarian cancer.Which is closely related to the process and metastasis of the tumor.Although recent progress has been made in the research of PDCD4 gene,the mechanism of PDCD4 down regulation and its down-regulation in tumor is not very clear.Our research team found that there are few reports of PDCD4 interacting proteins,especially in lung cancer.The interacting proteins may be related to function,mechanism and the occurrence and development of diseases.Therefore,we selected the interaction of PDCD4 protein as the research object,confirmed the interaction of PDCD4 with protein clear through the experiment,to investigate the expression of PDCD4 interacting protein in lung cancer and the correlation with PDCD4,which provides the beneficial help for the study of pathogenesis of non-small cell lung cancer.Purposes1.Screen for PDCD4 interacting proteins.2.Validate PDCD4 interacting protein.3.To clarify the expression of EIF3 H protein in non-small lung cancer cell and lung cancer tissues and its relationship with clinical parameters.4.Preliminary investigation of the relationship between PDCD4 and EIF3 H in non-small cell lung cancer cell and lung cancer.5.Screening and preliminary validation of EIF3 H interacting proteins.Methods1.Screening PDCD4 interacting protein The proteins that may interact with PDCD4 were screened by immunoprecipitation combined with mass spectrometry,retrieval results and subcellular localization.2.Validate PDCD4 interaction protein(1)The interaction between PDCD4 and EIF3 H was verified from both positive and negative sides by immunoprecipitation experiment.(2)Immunofluorescence was used to detect the localization of PDCD4 and EIF3 H.3.To clarify the expression of EIF3 H protein in non-small lung cancer cell and lung cancer tissues and its relationship with clinical parameters.(1)Expression of EIF3 H protein in non-small cell lung cancer cells by Western Blot.(2)The lung cancer tissue was detected by immunohistochemical,and the clinical parameters were analyzed by statistics.4.Preliminary investigation of the expression of PDCD4 and EIF3 H in non-small cell lung cancer cells and lung cancer.(1)The relationship between the mRNA level and the protein level of PDCD4 and EIF3 H in non small cell lung cancer cells by qPCR and Western Blot.(2)The expression of PDCD4 and EIF3 H in lung cancer tissues was detected by IHC.(3)The related factors affecting the prognosis of lung cancer patients by single factor and multiple factor regression analysis.5.Screening and preliminary verification of EIF3 H interaction protein(1)The EIF3 H interaction protein was screened by immunoprecipitation combined with mass spectrometry,retrieval results and subcellular localization.(2)It was positively verified by immunoprecipitation experiments whether NUDT21 is an interacting protein of EIF3 H.Results1.Screening possible PDCD4 interacting proteins: EIF3H;2.EIF3 H is a PDCD4 interacting protein;3.Compared with 16 HBE of human immortalized bronchial epithelial cells,EIF3 H was highly expressed in H1299,H460 and H1975 cells.There was no significant difference in EIF3 H expression between SPCA1 and A549 cells.Compared with adjacent cancer,EIF3 H was highly expressed in lung cancer tissue,and the difference was statistically significant;4.In non-small cell lung cancer cells,PDCD4 down-regulates the expression of EIF3 H mRNA and protein levels;EIF3H up-regulates the expression of PDCD4 mRNA levels,but PDCD4 protein levels did not change significantly.In lung cancer,PDCD4 was mainly low expression and EIF3 H was highly expressed.Single-factor regression analysis showed that clinical staging and N staging were related to the prognosis of patients with lung cancer.Multivariate regression analysis revealed that N staging could be used as an independent prognostic factor.5.NUDT11 may interact with EIF3 H protein.Conclusions:1.EIF3 H interacts with PDCD4.2.EIF3 H is highly expressed in lung cancer tissues and may play a certain biological function in lung cancer.3.PDCD4 may regulate the expression of EIF3 H at the level of transcription and translation;EIF3H may only affect the expression of PDCD4 at the transcriptional level.There was a significant negative correlation between the expression of PDCD4 and EIF3 H in lung cancer tissues.N staging can be used as an independent prognostic factor and may be related to the prognosis of lung cancer patients.4.NUDT11 may interact with EIF3 H protein. |
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