Magic™ Anti-IGFBP5 monoclonal antibody, clone C2697N

Increased histone H3R2me2as level is an independent prognostic factor in gastric cancer, which corresponds to PRMT6 overexpression. PRMT6-H3R2me2as pathway is activated and transcriptionally represses expression of tumor suppressor PCDH7, resulting in increased invasiveness of gastric cancer cells. Abstract Histone modification plays important molecular roles in development and progression of cancers. Dysregulation of histone H3 arginine (R) methylation is still unknown in primary cancer, including gastric cancer (GC). Although PRMT6 contributes to asymmetric dimethylation at H3R2 (H3R2me2as) in cancer cells, its molecular functions are poorly understood in GC. In this study, we assessed H3R2me2as and PRMT6 expression levels in 133 primary GC tissues by immunohistochemistry. Increased H3R2me2as was found in 68 GC (51.1%) cases and independently related to poor prognosis. PRMT6 was overexpressed in 70 GC (52.6%) and strongly correlated with the global H3R2me2as levels (P < 0.001). By analyzing biological functions of PRMT6 in GC cell lines by lentivirus-based systems, PRMT6 overexpression enhanced global H3R2me2as and invasiveness in vitro, while PRMT6 knockout (PRMT6-KO) suppressed these effects and tumorigenicity in vivo. ChIP and microarray assays demonstrated that PRMT6-KO GC cells decreased the enrichments of H3R2me2as at the promoter regions of PCDH7, SCD and IGFBP5, resulting in upregulation of their gene expression. PRMT6 was recruited to the promoter regions of PCDH7 and SCD in the PRMT6-overexpressed cells. Knockdown of tumor suppressor PCDH7 in the PRMT6-KO GC cells elevated cell migration and invasion. PRMT6 expression inversely correlated with PCDH7 expression in primary GC (P = 0.021). Collectively, our findings strongly indicate that H3R2me2as is a strong prognostic indicator of GC patients, and PRMT6-overexpressing GC cells may acquire invasiveness through direct transcriptional inhibition of PCDH7 by increasing H3R2me2as level. Thus, inhibition of the PRMT6-H3R2me2as pathway could be a promising new therapeutic strategy in GC.

Mammary epithelial cell (MEC) growth is reduced in continuously milked (CM) mammary glands, and administration of a mammogenic compound such as prostaglandin E-2 (PGE(2)) at parturition might improve MEC growth in CM tissue. The objectives were to 1) compare MEC turnover, ultrastructure, and gene expression in CM and involuting mammary tissue, and 2) evaluate the effects of CM and intramammary infusion of PGE(2) on early lactation MEC turnover, ultrastructure, mammary gene expression, milk yield, and composition. First- and second-lactation cows (n = 8) were used in a half-udder model, in which one-half was dry for 60 d (CTL) and the other was CM. Udder halves (n = 16) were assigned to a postpartum (PP) treatment of PGE2 (+ PGE(2); 875 mu g/10 mL of medium-chain triglyceride oil) or no PGE2 (-PGE(2)) treatment at parturition and at 72 h PP. Biopsies of CM and CTL quarters were obtained during milk stasis (MS) of the CTL half at 3 and 7 d after dry-off of the CTL half (3d-MS; 7d-MS) and postpartum (PP) at 2 and 4 d (2d-PP; 4d-PP). Milk yield was reduced (P < 0.01) in CM udder halves compared with CTL halves (13.2 vs. 22.1 kg/d), but reductions were less in second-lactation cows. The apoptotic index was greater (P < 0.05) in CTL glands than in CM glands (3d-MS, 0.52 vs. 0.11% and 7d-MS, 0.24 vs. 0.12, respectively). Proliferation of MEC was unchanged at 3d-MS, but was increased (P = 0.01) in CTL halves at 7d-MS compared with CM halves (3.10 vs. 0.93%). At 2d-PP, MEC proliferation was increased (P = 0.05) in CM halves compared with CTL halves (1.3 vs. 0.6%), but was unaffected by PGE(2) (P > 0.2). Apoptosis was elevated in early lactation regardless of treatment. Ultrastructure was unchanged by dry period length or PGE(2). In prepartum tissue, involution in CTL halves increased (P < 0.05) the expression of the proapoptotic genes Bcl-2-associated x protein (bax) and IGFBP5 and decreased (P < 0.05) alpha-lactalbumin expression compared with CM tissue. In PP mammary tissue, CTL halves expressed greater (P < 0.05) levels of aTPbinding cassette 1 (ABC1) and IGFBP5. Treatment with PGE(2) did not alter (P > 0.1) gene expression. The results confirm that CM reduced milk yield of cows with a mammary growth requirement. Reduced MEC turnover and milk yield were not alleviated by IMI of PGE(2), which indicates that peripartum PGE(2) concentrations in CM glands are not limiting mammary growth or milk synthesis.