The Correlation Between SATB1 Gene And Breast Cancer

Part I: The relationship between the SATB1 expression level and clinical pathological characteristics of breast cancerObjective: The immunohistochemistry method was used to detect the expression of SATB1 in breast cancer. To investigate its relationship with patients clinical pathological characteristics.Methods: 292 cases of breast cancer accepted surgery in guangxi medical university cancer hospital from January 2001 to December 2003 were enrolled in this study. Immunohistochemistry method was applied to detect the SATB1 expression on paraffin-embedded tissues of selected cases. To analysis the correlation between the expression level of SATB1 and clinical pathological characteristics through chi-square testResults: 208 analysable cases of breast cancer paraffin-embedded tissues were enrolled in this study finally. SATB1 staining was negative in 56 of 208(26.9%), and positive including 26 of"+"(12.5%), 80 of"++"(38.5%), 46 of "+++"(22.1%).There was no significant correlation of the level of SATB1 expression with ER,PR,HER2 and age(p>0.05).There was significant correlation of the level of SATB1 expression that greater than or equal to (++) with the lymph node metastasis and T stage.Conclusions: The level of SATB1 expression was not associated with hormone receptor status, HER2 status and patients age and significantly correlated with the lymph node metastasis and T stage.Part II: Explore the differentially expressed genes and its regulation relationships with SATB1 gene in breast cancer with bioinformaticsObjective: To screen the differentially expressed genes between breast cancer and adjacent tissues and its regulation relationships with SATB1 gene trough bioinformatics.Methods: The gene chip data of breast cancer were obtained from GEO(Gene Expression Omnibus)database and statistically analyzed using BRB-ArrayTools to identify the differentially expressed genes related to breast cancer with bioinformatics analysis using DAVID database on line Results: 70 differentially expressed genes were identified in breast cancer, including 28 up-regulated genes and 42 down-regulated genes. These genes were associated with the transcriptional regulation,cell signal transduction,cell cycle,cell adhesion,cell apoptosis etc. Collagen gene family is the main part among the up-regulated genes co-expressed with SATB1.There was no mutual regulation between SATB1 gene and its up-regulated genes.Conclusions: Bioinformatics analysis can successfully find gene cluster co-expressed with SATB1, which provide the premise for further reveal the correlation with its co-expressed genes.Part III: Cloning of SATB1 and construction of its eukaryotic expression vectorObjective: To investigate the function and relationship with other gene of SATB1, the ORF of SATB1 gene from the genomics was cloned, its eukaryotic expression vector pEGFP-N1-SATB1 was constructed.Methods: SATB1 cDNA library (pCMV6-XL6-SATB1) as a PCR template, PCR was performed with a pair of SATB1 specific primer. The PCR products were inserted into pGEM-T-Easy, and the resulted plasmid was transformed into DH5a.The positive clone was selected and sequenced. Then the gene that was confirmed by DNA sequencing was sucloned into pEGFP-N1 to construct eukaryotic expression vector.Results: The result of sequencing analysis is consistent with that of the references published (BC001744.1). Furthermore, a recombinant plasmid pEGFP-N1-SATB1 for eukaryotic expression was also obtained after subcloning.Conclusions: The SATB1 gene has been cloned successfully, and its eukaryotic expression vector with report gene a enhance green fluorescent protein (EGFP) gene has been obtained, The results have provide basis for further studies on the function of SATB1 and the correlation with other genes.