Peter Selvanayagam

Desquamin is an epidermal ribonuclease

Desquamin is a glycoprotein that we have isolated from the upper granular layer and the stratum corneum of human epidermis; it is not ordinarily expressed in submerged cultures, whose terminal differentiation stops short of formation of these layers. The exogenous addition of desquamin to human cultured keratinocytes extended their maturation, and hematoxylin staining indicated a loss of cell nuclei. For confirmation, cultured cells were lysed in situ, and the nuclei were incubated with desquamin for several days, then stained with hematoxylin. Damage to the nuclei was evident: the nuclear inclusions remained intact, while the surrounding basophilic nuclear matrix was degraded. Desquamin was then tested directly for nuclease activity. Ribonuclease activity was determined by incubating desquamin with human epidermal total RNA and monitoring the dose‐dependent disappearance of the 28S and 18S ribosomal RNA bands in an agarose/formaldehyde gel. On RNA‐containing zymogels, we confirmed the RNase activity to be specific to desquamin. Using synthetic RNA homopolymers, we found the active RNase domains to be limited to cytosine residues. On the contrary, DNA was not degraded by an analogous procedure, even after strand‐separation by denaturation. J. Cell. Biochem. 68:74–82, 1998.

Effects of Parathyroid Hormonal Peptides on the Gut1

Our previous studies have shown PTH to be an effective relaxant of smooth muscle throughout the mammalian tract. Recently, we found PTHrP to be equally as potent and effective on the gut as PTH, and we hypothesized that PTHrP, rather than PTH, might be the natural ligand for the gut receptors which mediate GI smooth muscle relaxation. To approach this question, we asked whether rat GI tissue expresses mRNA for PTHrP. Using selective reverse transcription and PCR we have found PTHrP mRNA in smooth muscle throughout the rat GI tract and in gastric and colonic mucosa as well. Our findings support the idea that PTHrP can be produced by GI tissues and that it may function there as an autocrine or paracrine factor. One of its actions may involve regulation of GI muscle tone.

Alteration of secretion of parathyroid hormone-related peptide and expression of its mRNA in a human hepatoma cell line (HEP G2) treated with agents that affect cell growth

Previously, using human hepatoma cells (HepG2), we found that immunoneutralization of secreted PTHrP increased cell growth. Here we asked whether PTHrP production was affected by agents that alter growth of Hep G2 cells. Immunoreactive PTHrP in medium and PTHrP mRNA expression were examined. Treatment of cells with 10 μM hydrocortisone or 1 ng/mL TGF-β1 for 72 h inhibited cell growth by 28±6 and 36±2% and increased PTHrP in medium by 128±10 and 525 ±27%, respectively. The increase in PTHrP produced by both agents was dose-and time-dependent, and the increased PTHrP was accompanied by dose-and time-dependent enhanced expression of PTHrP mRNA. In contrast, 10% fetal bovine serum (FBS) for 72 h increased cell growth by 38±6% (vs serum-free medium) and decreased PTHrP production by 49±4% whereas culture in high glucose (3–4g/L) increased cell growth by 43±1% (vs 1 g/L glucose) and decreased PTHrP by 55±0.4%. Inhibition of PTHrP by both FBS and glucose was dose-dependent; FBS also inhibited PTHrP mRNA. The results show that increased cell growth was associated with decreased PTHrP production, while decreased growth was accompanied by increased PTHrP production. The findings imply that PTHrP may help mediate growth effects of these agents on Hep G2 cells.