Goat anti-Human Stefin B polyclonal antibody

The prognosis for pancreatic ductal adenocarcinoma (PDAC) remains poor despite decades of effort. The abundant extracellular matrix (ECM) in PDAC comprises a major fraction of the tumor mass and plays various roles in promoting resistance to therapies. However, nonselective depletion of ECM has led to poor patient outcomes. Consistent with that observation, we previously showed that individual matrisome proteins derived from stromal cells correlate with either long or short patient survival. In marked contrast, those derived from cancer cells correlate strongly with poor survival. Here, we studied three cancer cell-derived matrisome proteins that are significantly overrepresented during PDAC progression, AGRN (agrin), SER-PINB5 (serine protease inhibitor B5), and CSTB (cystatin B). Using both overexpression and knockdown experiments, we demonstrate that all three are promoters of PDAC metastasis. Furthermore, these proteins operate at different metastatic steps. AGRN promoted epithelial-to-mesenchymal transition in primary tumors, whereas SERPINB5 and CSTB enhanced late steps in the metastatic cascade by elevating invadopodia formation and in vivo extravasation. All three genes were associated with a poor prognosis in human patients and high levels of SERPINB5, secreted by cancer cells and deposited in the ECM, correlated with poor patient prognosis. This study provides strong evidence that cancer cell-derived matrisome proteins can be causal in promoting tumorigenesis and metastasis and lead to poor patient survival. Therefore, compared with the bulk matrix, mostly made by stromal cells, precise interventions targeting cancer cell-derived matrisome proteins, such as AGRN, SERPINB5, and CSTB, may represent preferred potential therapeutic targets. Significance: This study provides insights into the biological roles of cancer cell-derived matrisome proteins in PDAC and supports the notion that these proteins are protumorigenic and better therapeutic targets.

The Unverricht-Lundborg type of progressive myoclonus epilepsy (EPM1) is an autosomal recessive disorder that is caused by the dysfunction of the cystatin B (CSTB) gene product. In the vast majority of affected cases, mRNA transcription is impaired by a biallelic expansion of a dodecamer repeat within the 5'-untranslated region of the respective gene. Since this minisatellite contains exclusively G and C nucleotides, direct PCR analysis of allele expansion is extremely difficult and error prone. To circumvent these problems, we have developed a PCR assay that is based on the deamination of the DNA prior to amplification. We have developed a method based on PCR after DNA deamination of the GC-rich repeat region, which improves the PCR condition to such an extent that we were not only able to reliably amplify expanded alleles of affected individuals (homozygotes and compound heterozygotes), but also the two alleles of full mutation carriers, whose analysis is particularly difficult because of PCR bias and heteroduplex formation between the two alleles. We used promoter, and repeat specific primer combinations to investigate whether dodecamer repeat expansion concurs with de novo methylation of the CSTB gene promoter in a similar fashion to other repeat expansion syndromes. We confirmed previous evidence obtained by HpaII digestion and Southern blot analysis that both the promoter and the repeat regions are unmethylated, in both healthy and affected individuals. Thus, in contrast to certain trinucleotide repeat expansion-associated diseases, such as fragile X syndrome (FRAXA) and myotonic dystrophy, methylation analyses can not be utilized for indirect diagnostic testing. (C) 2003 Wiley-Liss, Inc.