| IntroductionBreast carcinoma is one of major malignant tumors that endangered womens health and the leading cause of women death. Thus to elucidate its pathogenesis is of great significance for a better diagnosis and treatment.More and more data demonstrated carcinoma is a category cell cycle disease. Studies showed that the cell cycle was regulated by Cyc, CDKs and CKIs, and there are several key checkpoints between one phase and others. The checkpoint between G1 and S phase is a key point involved in the regulation of cell proliferation, through which the intracellular and extracellular signal can be transmitted and integrated to nucles. The dysplasia of this checkpoint is closely related to tumor development. Among the factors invoved in the regulation of cell cycle, the pathway composed of Rb,cyclinD1 ,CDK4/6 and p16 have been studied further and clearly, and is believed to be closely related to the checkpoint between G1and S phases. Normal cells entering S phase is positive regulated by cyclinD1 and negative regulated by p16 simul-tarously.Immunohistochemistry S-P method was used to detect the expression of p16 and cyclinD1 in human mammary atypical hyperplasia--a precancerous lesion of breast carcinoma, compared with benign hy-perplasia and breast carcinoma, to investigate their functional significance and relationship in the malignant progression of epithelium.Materials and methodsAll 70 tissue samples were taken from Pathology Department of Shenyang Military Region General Hospital at random. We selected 30 cases of mammary hyperplasia, including 6 benign hyperplasia, 11 mild atypical hyperplasia, 12 medium atypical hyperplasia, 21 severe atypical hyperplasia and 20 cases of breast carcinoma (8 intraductal carcinoma and 12 invasive ductal cacinoma). For immunostainy, streptavidin peroxidase complex method was used, p16, cyclinD1 were used as first antibody, the S-P kit was purchased from fuzhou maixin biocompany. The sections were dewaxed, and then treated with 3% hydrogen peroxide in methanol for 10 minutes to quench the endogenous peroxidase activity. The sections were incubated for 30 minutes with normal non-immuneserum to eliminate nonspecific staining. The sections were incubated overnight at 4 with primary antibody. This was followed by incubation with biotin-labelled secondary antibody for 30 minutes and streptavidin-peroxidase complex for 30 minutes. The color was developed with diaminobenzidine and the sections were counterstained with harmatoxylin. Negative control was carried out by replacing the primary with PBS. The extent of staining was scored as (-) indicating < 10% of tumor area stained; ( +)10% -30% stained; ( + + ) 30% -60% stained; ( + + + ) > 60% stained. The statistical methods were according to x2 test and correlation analysis.Results1. The expression of cyclinD1 and p16The expression of p16 was localized in nucleus and cytoplasm, and cyclinD1 localized in nucleus, brownish yellow granule could be observed. The expression of p16 were the highest in benign hyperplasia (83. 3% ) (mild 81. 8% , medium 75. 0% , severe 66. 7% , cancer 55. 0% ). That the expression of p16 in benign hyperplasia ,atypical hyperplasia is obviously higher than breast caninoma and statistics showed the difference was obvious (P <0. 05). The expression of cy-clinD, were negative in benign hyperplasia and mild atypical hyperplasia and with the hyperplasia degree of ductal epithelium, the expression of eyeing increased, especially obvious in medium-severe atypical ductal hyperplasia. The difference between severe atypical hyperplasia ,.carcinoma and benign hyperplasia,mild atypical hyperplais was obvious (P <0. 05) , while the difference between medium-severe a-typical and carcinoma was no significant ( P >0. 05).2. The relationship between the expression of p16 and cyclinD1 Statistics showed that the expression of p16 was correlated withthe expression of cyclinD1 ( P < 0. 05 ) , correlation coefficient was -0.567.Disussion1. The relationship between the a...
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