| IntroductionBreast cancer is one of the most common malignancies affecting women and the leading cause of women cancer mortality. A good deal of studies showed the development of breast carcinoma was associated with the atypical ductal hyperplasia. Presently several observations suggested a prominent role for the mitogen - activated protein kinase (MAPK) regulatory network in the development of breast carcinoma. MAPK cascade pathway can converge extracellular signals and play an important role in signal transduction.Extracellular signal - regulated kinases ( ERK) , is a firstly discovered member of MAPK family. It is a kind of highly conserved and ubiquitously expressed serine/threonine kinase activated either by growth factors binding to tyrosine - kinase - linked receptor or by lig-ands binding to G - protein - linked receptors. ERK participates some physiological progressions such as cell growth, development, mitosis and functional synchronism, and plays a role in the pathological progression of maliganant transformation of cell. Phospho - ERK, or phosphorylated activated ERK, may translocate into nucleus to regulate the expression of certain genes through phosphorylation on transcription factors. It had been studied that activation of ERK/MAP kinase is necessary and sufficient for PC 12 differentiation and for transformation of NIH3T3 cells, and is indispensable for tumor formation. It was not until 1997 that Sivaraman found that ERK/MAPK is overex-pressed and hyperactivated in breast carcinoma. And it is the first evidence showing ERK/MAPK contribute to the transformation of tumor phenotype. Since then studies gradually increased on ERK/MAPK in other epithelium derived neoplasms.S - P method of immunohistochemistry was used to detect the protein expression of ERK2/p - ERK in human mammary hyperplasia and carcinoma, assessing the potential roles of ERK/MAPK in malignant progression of epithelial cells. There we used a newly developed antibody specific for activated ERK/MAPK (phospho -ERK) to investigate the activation of ERK and observe its spatial patterns in various breast diseases. The assessment of the level of activated ERK with phosphorylated specific antibodies hasn ' t been reported much in human tumor tissues, and especially it hadn' t been reported in human breast carcinoma.Materials and MethodsAll 67 samples were obtained from the Pathology Department of the First Affiliated Hospital of China Medical University and the Second. They were female, age ranging from 21 to 73 years. They were classified with the Histologically Standard of Breast Hyperplasia and Neoplasms recommended by the Editorial Broad of Chinese Journal of Pathology.Immunohistochemistry S - P method was used to investigate the protein expression of ERK2 and p - ERK in 43 cases of mammary hyperplasia (including 8 benign hyperplasia, 10 mild atypical hyperplasia, 12 medium atypical hyperplasia, and 13 severe atypical hyperplasia ) as well as 24 cases of breast carcinoma (11 intraductal carcinoma and 13 invasive ductal carcinoma). Antigen unmasking were per-formed for 25 minutes with complex - enzyme at 37 C. ERK2( Santa Cruz Biotechnology, cat# sc - 154) and p -ERK(Santa Cruz Biotechnology, cat# sc -7383) were incubated for 120 minutes at a dilution of 1:100 and for 90 minutes at a dilution of 1:200, respectively. Between each steps washed in PBS three times for 5 minutes, with the exception of removing the serum using suction. Visulization of antibody binding was the DAB method. PBS replaced the primary antibod-y as a negative control.Tissue sections stained with ERK2 were quantitated subjectively using the following scale ( 0: none, 1 + : weak, 2 + + : strong). Tissue sections stained with p - ERK were quantitated subjectively using the following scale( 0; none; 1 + ; positive cells percentage < 10% ; 2 + : positive cells percentage >10% ).Statistical analysis: The data analyses were performed using SPSS for Windows 10. 0 with a significant level of P <0. 05.Results1. The protein expression of ERK...
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