| Objective It was found that CCDC170 was highly correlated with ESR1,and distinct in different pathotyping.And it affected the patient's prognosis.After that,we used MCF-7 cells to over expression CCDC170 24 h and 48 h,and conducted m RNA scan.Pathway analysis was used to find related genes.Among them,IRE1 is the key gene of the UPR,and protein IRE1? can shear XBP1 m RNA,initiate UPR,and regulate cell survival.XBP1 can play the role of transcription factors by combining with the X box sequence of the target gene promoter region.XBP1 can be combined with ER?,and improves the transcriptional activity of ER?.The purpose of this study was to investigate the relationship between IRE1?,CCDC170,ER?,XBP1 and the clinical parameters;furthermore,to explore the possible mutual regulation between them.Methods IRE1 was found through microarray technology,IPA analysis.The m RNA expression data were from TCGA.100 cases of breast invasive ductal carcinoma from November 2008 to April 2009 were selected.Immunohistochemical staining was used to detect the protein expression and location.SPSS19.0 was used to analyze data.Use Graphpadprism5.0 for drawing,all the statistics were bilateral probability test,and P < 0.05 had statistical significance.Immunofluorescence technique and Western Blot showed the change of IRE1?,XBP1 and ER? after overexpression of CCDC170.And immunofluorescence observed the changes of cytoskeletal proteins.Results 1.IPA analysis found over expression CCDC170 24 h and 48 h,the m RNA expression of IRE1 were all increased.2.At the level of m RNA,IRE1,ER? and XBP1 was positively correlated with CCDC170(P < 0.05)from the data of TCGA.3.CCDC170 protein was mainly located in the cytoplasm,and its expression of(-),(+),(+ +)and(+ + +)were13,37,35 and 15 cases,respectively.IRE1? protein was located in cytoplasm,and its expression of(-),(+),(+ +)and(+ + +)were 16,57,19 and 8 cases,respectively.XBP1 protein was mainly found in the nucleus,and its expression of(-),(+),(+ +)and(+ + +)were 7,35,33 and 25 cases.4.CCDC170 was positively correlated with ER?(r = 0.306,P = 0.002),and negatively with Her-2(r = 0.202,P = 0.044).IRE1? was positively related with Her-2(r = 0.315,P = 0.001).XBP1 was positively associated with lymph node metastasis(r = 0.248,P = 0.013)and ER?(r = 0.280,P = 0.005).5.CCDC170,XBP1,and lymph node status,TNM affect the overall survival significantly.The tumor size,lymph node metastasis,and TNM was associated with disease-free survival in Log-rank test analysis;but in Multivariate Cox proportional hazards regression analysis,only TNM remains significant for overall and disease free survival,and CCDC170 affectd overall survival merely.6.Western Blotting found over expression CCDC170 for 24 h,the protein IRE1? increased,and all protein mentioned above increased obviously at 48 h.The immunofluorescence show similar results,the change of XBP1 is more obvious,and cell cytoskeleton was influenced.Conclusions 1.The expression of CCDC170 was highly associated with IRE1?,ER? and XBP1 at the level of m RNA from the data of TCGA.2.CCDC170 and IRE1? are in cytoplasm and XBP1 in nucleus.CCDC170 expression was correlated with ER?,XBP1 and Her-2.IRE1? was positively rrelated with Her-2.XBP1 was correlated with lymph node and ER?,suggesting that they may be related to the occurrence and development of breast cancer.3.The level of CCDC170 had effect on OS significantly.Patients with high of CCDC170 had good prognosis.But for XBP1,the result was opposite.Therefore,more reliable prognostic implications will be obtained by expanding sample size.4.CCDC170 can change cell cytoskeleton and promote the expression of IRE1?,ER? and XBP1,suggesting that there may be potential regulatory mechanisms between CCDC170,IRE1?,ER? and XBP1. |
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