6273

S100A2

CAN19|S100L;S100 calcium binding protein A2;S100A2;S100 calcium binding protein A2

S100 calcium-binding protein A2|protein S100-A2

1q21

protein-coding

BACKGROUND: Our cDNA microarray analysis of squamous cell carcinoma of the head and neck (SCCHN) previously identified that S100A2 was down-regulated in highly metastatic 686LN-M3s cell lines established through in vivo selection using a metastatic xenograft mouse model. S100A2, a putative tumor suppressor, has been found to be down-regulated in several types of primary tumor as compared with the normal tissue. Only a few reports have explored its expression status and function in metastasis. METHODS: To further confirm down-regulation of S100A2 in human metastasis, we examined S100A2 expression using immunohistochemical analysis of paraffin-embedded SCCHN tissues. The samples included primary SCCHN tumors (Tu-1) and involved lymph nodes (Met-1) from the same patients, and primary tumors in node-negative patients (Tu-2). RESULTS: Most of these tumors expressed S100A2 but lymph node metastases showed a pattern of reduced staining for S100A2 compared with primary tumors. A similar expression pattern of S100A2 was also observed in several SCCHN cell lines by reverse transcription-polymerase chain reaction (RT-PCR) and immunoblotting. Particularly, S100A2 expression was lower in 686LN than Tu686 and hardly detectable in the metastatic derivatives 686LN-M3s. Further study of S100A2 promoter showed higher methylation intensity in these metastatic derivatives than in Tu686 and 686LN. CONCLUSIONS: S100A2 was down-regulated in lymph node metastasis of SCCHN, suggesting that instead of being a putative tumor suppressor, S100A2 may play a role in the metastasis of SCCHN.

S100A2 is generally found expressed in the epidermis and was recently shown to play a crucial role in the differentiation of keratinocytes. Also known as CaN19, S100A2 was identified as a potential tumor suppressor. expression of S100A2 is upregulated by p53. The proteins p63 and p73 are related to p53 and are expressed as several splice variants with partially overlapping tasks but also functions different from p53. It had been shown that p63 proteins with mutations in their DNA-binding domain cause severe phenotypes in man as autosomal dominantly inherited disease including EEC, AEC, SHFM, LMS and ADULT syndromes. Here we show that S100A2 is a transcriptional target of p63/p73 family members, particularly the p63 splice variant TAp63gamma. The regulation is mediated by a novel transcriptional element in the S100A2 promoter which is bound by TAp63gamma but not by p53. Mutant p63 proteins derived from EEC and ADULT syndrome patients cannot activate S100A2 transcription whereas SHFM-related mutants still can stimulate the S100A2 promoter. Consistent with a function in tumor suppression S100A2 expression is stimulated upon DNA damage. After doxorubicin treatment p63gamma proteins are recruited to the S100A2 promoter in vivo. This may indicate a function of the p63-dependent S100A2 regulation in tumor suppression.

PURPOSE: In primary squamous cell carcinoma of the larynx (LSCC), Ca(2+) binding S100A2 protein underexpression was already found to be associated with poor tumour differentiation and shorter overall survival. In the present work, the role of S100A2 protein expression in the prediction of regional metastasis-free survival (MFS) was investigated to guide neck management in LSCC. EXPERIMENTAL DESIGN: Specimens of LSCC from 62 consecutive untreated patients were examined for S100A2 content by immunocytochemistry; the patients were followed up for a median of 44 months (range 2-90 months) after initial surgical resection. MFS was calculated from the date of first surgery to that of regional neck node recurrence. RESULTS: S100A2 was detected in 18 of 19 (95%) low-grade tumours and in 22 of 43 (51%) high-grade tumours. The 5-year regional MFS was 81% for patients with S100A2-positive tumours and 55% for patients with S100A2-negative tumours. By multivariate analysis, the S100A2 status appeared to be a significant independent predictive factor for MFS (p = .02). CONCLUSIONS: Our results suggest that the assessment of S100A2 status at diagnosis may identify a subset of LSCC patients highly susceptible to neck node metastases and may thus help define therapy accordingly.

The calcium-binding protein S100A2 is expressed in normal breast tissue but downregulated during breast cancer progression. Hence it was previously identified as a candidate tumor suppressor gene. In this report, we investigated the molecular basis of S100A2 gene expression in normal and tumorigenic human breast epithelial cells. We cloned the gene coding for S100A2 including its 5 flanking region. To locate positively or negatively acting elements responsible for transcriptional regulation, promoter deletion studies were performed. Results from these experiments demonstrate that an enhancer element is located 1.2 kb upstream of the transcription start site. This element contains two AP1-like binding sites suggesting that transcriptional activation of S100A2 might be mediated by immediate early genes. Interestingly, the enhancer stimulates transcription in both normal and tumorigenic cells, indicating that repression of endogenous S100A2 transcription in tumorigenic cells might lie at an epigenetic level. Indeed, the proximal promoter region was found, by genomic sequencing, to be unmethylated in normal but hypermethylated in tumorigenic cells. Hypermethylation of the promoter at the same CpG sites was also found in a breast cancer biopsy. In addition, site specific in vitro methylation led to reduced expression of the S100A2 gene in normal cells. These experiments provide strong evidence that S100A2 repression in tumor cells is mediated by site-specific methylation. Since transcription of a number of known tumor suppressor genes is also repressed by methylation, our observation is consistent with the suggestion that S100A2 might have a tumor suppressor function.