Collagen type I-induced Smad-interacting protein 1 expression downregulates E-cadherin in pancreatic cancer.

Pancreatic cancer is a devastating disease with poor prognosis. Production of large quantities of extracellular matrix and early metastasis are characteristics of this disease. One important step in the development of various cancers is the loss of E-cadherin gene expression or inactivation of E-cadherin mediated cell-cell adhesion. It has been shown that collagen type I promotes downregulation of E-cadherin expression, which correlates with enhanced cell migration and invasiveness. In this context, we elucidated the role of Smad-interacting protein 1 (SIP1), which has been discussed as a negative regulator of E-cadherin gene expression. We demonstrate that SIP1 upregulation shows an inverse relationship with E-cadherin in advanced pancreatic tumour stages. In Panc-1 cells, SIP1 expression can be induced by exposure to collagen type I in a src-dependent manner. In addition, overexpression of SIP1 reduces E-cadherin mRNA and protein levels. Taken together, these results suggest that SIP1 is involved in the progression of pancreatic cancer and plays a role in mediating signal transduction from collagen type I to downregulate E-cadherin expression.

Laser-mediated microdissection of paraffin sections from Xenopus embryos allows detection of tissue-specific expressed mRNAs.

One of the key end points for understanding the molecular basis of embryogenesis is the analysis of spatiotemporal patterns of gene expression. Methodical limitations due to low mRNA levels often prevent a tissue-specific resolution. In this study, we developed an improved laser microdissection technique and RT-PCR that allows marker gene detection in small tissue areas from sections of formalin-fixed paraffin-embedded Xenopus embryos. Tissue pieces were isolated by laser microbeam microdissection and captured by laser pressure catapulting. Neither laser treatment nor conventional histological or immunochemical staining impaired subsequent RNA analysis. Transcripts of tissue-specific marker genes such as endodermin (endoderm), epidermal cytokeratin (epidermal ectoderm), N-CAM (neural tube), myoD (somites), and sonic hedgehog (floor plate) were amplified by nested RT-PCR analysis from small areas of single sections.

Molecular characterization and expression pattern of XY body-associated protein XY40 of the rat.

The XY body is a structure formed by the partially synapsed chromosomes X and Y that is located at the nuclear periphery of mammalian pachytene spermatocytes. In contrast to the autosomal bivalents of the same nucleus, the XY body is characterized by its differential chromatin condensation and transcriptional inactivity. In order to shed some light on the biological significance of these differences we have been characterizing XY body-associated proteins. We present here the cDNA sequence and expression pattern of XY40, a protein that is associated with the axial elements of the XY body. RNA blot analysis revealed that during spermatogenesis the transcript that encodes protein XY40 was highly enriched in pachytene spermatocytes. This transcript was also detectable in brain and, to a lesser extent, in liver and kidney. Although the signal in brain was as strong as in spermatocytes, protein XY40 could be detected only in the latter. The nucleic acid sequence reveals that XY40 is a novel protein with a few similarities to already known nucleotide sequences. Among these similarities the most interesting is a box that is shared by the 3' untranslated region of XY40 and the 5' untranslated region of Munc-18c, a member of a protein family involved in synaptic vesicle exocytosis. Since the transcripts of both XY40 and Munc18-c show a similar expression pattern, it is tempting to speculate that this common sequence is involved in translation regulation.