There is considerable need for reliable prognostic markers to guide clinicians in management decisions for the breast cancer. The cell cycle is governed by a family of cyclin-dependent kinases(Cdks), regulated by associated cyclin p27, a cyclin dependent kinase inhibitors, regulates progression from GI into S phase by inhibiting cyclin/cdks complex. This study was performed to evaluate the association between p27 expression, determined by immunohistochemical stain, and various histopathologic features in breast cancer. It was also determined whether p27 expression had the significance as the prognostic factorin the breast cancer patients. 45 patients who got the relatively good preserved paraffin blocks among the 100 patients was chosen for immunohistochemical staining against p27, cyclin E, c-erbB2, cathepsin D and p53 and reexamined their unclear and histological grades from Jan. 1989 through Dec. 1992. As a results, the patients with negative expression of p27 had more metastatic axillary nodes than those with positive expression. (5.87+/-1.87 vs 1.14+/-0.54, p=0.021) p27 negative group got worse nuclear grade than p27 negative group but beyond statistical significance. (48.4% vs 28.6%, p=0.281) On univariate analysis, primary tumor size, status of axillary nodes and the expression of p27 were the significant prognostic factors affecting overall survival rates. In particular, p27 positive group had better outcomes on 5 years survival rate than p27 negative group. (92.31+/-7.39% vs 73.33+/-8.07%, p=0.0441) but didn't affect the disease free survival with statistical significance. On multivariate analysis, the primary tumor size and axillary node were significant prognostic factors on overall and disease free survival. In conclusion, despite relativeiy small size of this study group, considering that p27 negative group got the more metastatic node and worse overall survival, 27 expression might be a novel prognostic indicator in the breast cancer.
Breast Neoplasms* ; Breast* ; Cathepsin D ; Cell Cycle ; Cyclin E* ; Cyclins* ; Disease-Free Survival ; Humans* ; Multivariate Analysis ; Paraffin ; Phosphotransferases ; S Phase ; Survival Rate

OBJECTIVETo screen the differentially expressing genes between silicotic lung tissue and normal lung tissue, to identify the differentially expressing genes of matrix metalloproteinase-12 (MMP-12) and Cathepsin E and to explore the roles of those genes in silicosis development.

METHODSThirty male SD rats were divided randomly into two groups: control group (6 rats) and exposure group (24 rats) which was exposed to SiO2 by intra-tracheal perfusion. On the 30 th, 60 th and 90 th days after exposure, 8 rats in model group and 2 rats in control group were executed and the lung tissues were obtained. The morphologic changes of lung tissues were observed with HE staining and VG staining under a light microscope. The gene microarrays were used to identify differentially expressing genes of lung tissues in rats exposed to SiO2 for 60 days. Two significantly up-regulated genes, MMP-12 and Cathepsin E, were validated using RT-PCR, immunohistochemistry and Western Blot assay.

RESULTSA total of 338 differentially expressing genes were identified from the 26 962 genes between silicotic rats and normal rats, including 267 up-regulated genes and 71 down-regulated genes. The results of RT-PCR showed that in the lung tissues of exposure group on the 30 th, 60 th and 90 th days, the mRNA expression levels of MMP-12 were 4.306, 5.338, 6.713 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.434, 2.974, 3.889 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the mRNA expression levels of MMP-12 were 1.435, 1.746, 2.069 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.372, 1.663, 2.103 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the expression levels of MMP-12 protein were 1.214, 1.531, 1.959 times higher than those in the control group, the expression levels of Cathepsin E protein were 1.262, 1.828, 1.907 times higher than those in the control group, respectively. Compared with the control group, the mRNA and protein expression levels of MMP-12 and Cathepsin E in lung tissues of exposure group were significantly up-regulated (P < 0.05).

CONCLUSIONThe differentially expressing genes in rat lung tissues screened by gene chip were validated, which suggested that a complex gene regulatory network may be contributed to occurrence of silicosis. MMP-12 and Cathepsin E genes may be involved in the development of silicotic pulmonary fibrosis by degrading the basement membrane of alveolar wall and participating in the immune response.

Animals ; Cathepsin E ; genetics ; metabolism ; Gene Expression ; Lung ; metabolism ; Male ; Matrix Metalloproteinase 12 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Silicosis ; genetics ; metabolism

PURPOSE: The ketogenic diet has long been used to treat epilepsy, but its mechanism is not yet clearly understood. To explore the potential mechanism, we analyzed the changes in gene expression induced by the ketogenic diet in the rat kainic acid (KA) epilepsy model. MATERIALS AND METHODS: KA-administered rats were fed the ketogenic diet or a normal diet for 4 weeks, and microarray analysis was performed with their brain tissues. The effects of the ketogenic diet on cathepsin E messenger ribonucleic acid (mRNA) expression were analyzed in KA-administered and normal saline-administered groups with semi-quantitative and real-time reverse transcription polymerase chain reaction (RT-PCR). Brain tissues were dissected into 8 regions to compare differential effects of the ketogenic diet on cathepsin E mRNA expression. Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks. RESULTS: The microarray data and subsequent RT-PCR experiments showed that KA increased the mRNA expression of cathepsin E, known to be related to neuronal cell death, in most brain areas except the brain stem, and these increases of cathepsin E mRNA expression were suppressed by the ketogenic diet. The expression of cathepsin E mRNA in the control group, however, was not significantly affected by the ketogenic diet. The change in cathepsin E mRNA expression was greatest in the hippocampus. The protein level of cathepsin E in the hippocampus of KA-administered rat was elevated in immunohistochemistry and the ketogenic diet suppressed this increase. CONCLUSION: Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.
3-Hydroxybutyric Acid/blood ; Animals ; Cathepsin E/genetics/*metabolism ; Enzyme Activators/pharmacology ; *Gene Expression Regulation, Enzymologic/drug effects ; Hippocampus/*drug effects/*metabolism ; Immunohistochemistry ; Kainic Acid/*pharmacology ; *Ketogenic Diet ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction