5655

KLK10

NES1|PRSSL1;kallikrein-related peptidase 10;KLK10;kallikrein-related peptidase 10

breast normal epithelial cell associated serine protease|kallikrein 10 protein 1|kallikrein 10 protein 12|kallikrein 10 protein 2|kallikrein 10 protein 3|kallikrein 10 protein 4|kallikrein 10 protein 7|kallikrein 10 protein 8|kallikrein 10 protein 9|kalli

19q13

protein-coding

The normal epithelial cell-specific 1 (NES1) gene (official name kallikrein gene 10; KLK10) is a new member of the expanding human kallikrein gene family and encodes for a secreted serine protease. Experimental evidence suggests that NES1 controls normal cell growth and may function as a tumour suppressor. NES1 is down-regulated during breast cancer progression. The NES1 gene is highly expressed in testicular as well as in other tissues. In this study, we investigated the expression level of the NES1 gene in cancerous and normal testicular tissues with reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. In all 14 primary testicular germ-cell tumours examined, the NES1 gene expression was markedly reduced compared to adjacent (paired) normal tissues. We further examined 6 randomly selected primary germ-cell tumours and 8 normal tissues (obtained from different individuals). We confirmed the differential expression of the NES1 gene in germ-cell tumours (GCT) and pre-malignant carcinoma in situ (CIS). Our findings suggest that NES1 may act as a tumour suppressor and may play a role in the pathogenesis and progression of this malignancy.

PURPOSE: The loss of expression of NES1, a novel putative tumor suppressor gene, is an early marker of breast tumorigenesis. NES1 is expressed in normal breast tissue and ductal hyperplasia but is absent or markedly diminished in invasive cancer. In cases of ductal carcinoma in situ (DCIS), NES1 expression has been shown previously to be present in approximately 50% of specimens. This study examined the expression level of NES1 in diagnostic biopsy samples found to contain pure DCIS. These data were then correlated with the pathologic findings found at definitive local surgery. METHODS AND MATERIALS: Twenty-nine cases with initial biopsy showing DCIS without invasive carcinoma followed by subsequent reexcision were discovered and archived. Formalin-fixed tissue specimens were obtained for analysis. Each biopsy specimen was subjected to hematoxylin-eosin staining and reviewed by two pathologists to confirm the diagnosis of pure DCIS. NES1 cDNA (1069 bp), including 238 bp of 5 and 3 untranslated region and the entire protein-coding region, was cloned into a vector. To generate the antisense and sense RNA probes, the plasmid was linearized and the transcription reaction was carried out with polymerases T7 and T3, respectively. The detection of in situ hybridization probes was performed using an mRNAlocator-Biotin Kit. Staining was characterized as negative (0/1+) or positive (2+/3+). Subsequent to an initial biopsy diagnosis of DCIS, all cases had a definitive surgical procedure. Detailed sectioning of the resultant tissue was performed and subjected to hematoxylin-eosin staining to determine the presence or absence of invasive carcinoma. RESULTS: The initial diagnostic biopsy specimens showed that 17 of 17 high-grade, 3 of 7 intermediate-grade, and 3 of 5 low-grade DCIS specimens were negative for NES1 expression. Of the 6 cases of DCIS found to be positive for NES1 expression, none (0%) were subsequently found to have invasive carcinoma at definitive surgery. In contrast, the loss of NES1 expression in the initial diagnostic biopsy was associated with a 40% incidence of invasive carcinoma at definitive surgery. Additional stratification by nuclear grade showed invasive carcinoma in 5 (83%) of 6 NES1-negative, low- to intermediate-grade DCIS (p expression in DCIS identified at the diagnostic biopsy predicts for a high risk of invasive cancer in the definitive surgical specimen. The predictive value of NES1 expression appears to be particularly relevant for low- and intermediate-grade DCIS.

The c-Jun dimerization protein, JDP2, is a member of the AP-1 (activating protein-1) family of the basic leucine zipper transcription factors. JDP2 can bind 12-O-tetradecanoylphorbol-13-acetate (TPA)-responsive element and cAMP-responsive element DNA response elements, resulting in the inhibition of transcription. Although the role of AP-1 in cell proliferation and malignant transformation is well established, the role of JDP2 in this process is of subject to debate. On the one hand, JDP2 was shown to inhibit cyclin D transcription and promote differentiation of skeletal muscle and osteoclast cells. On the other hand, JDP2 was shown to partially transform chicken embryo fibroblast and was identified in a screen for oncogenes able to collaborate with the loss of p27kip cyclin-dependent inhibitor to induce lymphomas. Using cell transformation assays in NIH3T3 cells and injection of prostate cancer cell lines overexpressing JDP2 into severe combined immuno-deficient (SCID) mice, we show for the first time the potential role of JDP2 in inhibition of cell transformation and tumor suppression. The mechanism of tumor suppressor action of JDP2 can be partially explained by the generation of inhibitory AP-1 complexes via the increase of JunB, JunD, and Fra2 expression and decrease of c-Jun expression.

Previously (Liu et al, cancer Res., 56: 3371-3379, 1996), we isolated a novel serine protease-like gene--Normal Epithelial Cell Specific-1 (NES1)--that is expressed in normal mammary epithelial cells but is down-regulated in most breast cancer cell lines. Here, we demonstrate that stable expression of NES1 in the NES1-negative MDA-MB-231 breast cancer cell line suppressed the oncogenicity as revealed by inhibition of the anchorage-independent growth and tumor formation in nude mice. Fluorescence in situ hybridization localized the NES1 gene to chromosome 19q13.3, a region that contains genes for related proteases (including the prostate-specific antigen) and is rearranged in human cancers. Similar to breast cancer cell lines, prostate cancer cell lines also lacked NES1 mRNA and protein expression. Together, these results strongly suggest a tumor-suppressor role for NES1 in breast and prostate cancer.