MAML1 promotes ESCC aggressiveness through upregulation of EMT marker TWIST1.

BackgroundMastermind-like 1 (MAML1) is the main transcriptional co-activator of Notch signaling pathway. It plays essential roles in several pathways including MEF2C, p53, Nf-кB and Wnt/β-catenin. TWIST1 is known as a regulator of epithelial mesenchymal transition (EMT), which is considered as a primary step in promotion of tumor cell metastasis. Since concomitant expression of these genes was observed in tumors, our aim in this study was to elucidate the linkage between MAML1 and TWIST1 co-overexpression in esophageal squamous cell carcinoma (ESCC).ResultsWhile MAML1 silencing significantly down-regulated TWIST1, its ectopic expression up-regulated TWIST1 expression in both mRNA and protein levels in KYSE-30 cells. Expression of mesenchymal markers was increased significantly after MAML1 and TWIST1 ectopic expression, while epithelial markers expression was significantly decreased after silencing of both genes. Concomitant protein expression of MAML1 and TWIST1 was significantly observed in ESCC patients. Enforced expression of TWIST1 had no impact on MAML1 gene expression in KYSE-30 cells.ConclusionThe results clearly suggest transcriptional regulation of TWIST1 by MAML1 transcription factor in ESCC cells KYSE-30. Since TWIST1 is known as an EMT inducing marker, our results may revealed the mastermind behind TWIST1 function and introduced MAML1 as an upstream master regulator of TWIST1 and EMT in KYSE-30 cells

Detection of heat-labile toxin in enterotoxigenic Escherichia coli using PCR-ELISA technique

Background and Objective: Enterotoxigenic Escherichia coli (ETEC) are the most common agent which causes diarrhea, worldwide. ETEC is colonized along the cells and then producing heat-labile (LT) and heat-stable enterotoxigenic which enter into intestinal epithelial cells and causes water and electrolyte loss from intestinal epithelial cells and eventually cause diarrhea.This study was done to detect the heat-labile toxin in Enterotoxigenic Escherichia coli using PCR-ELISA technique. Methods: In this descriptive study, DIG-labeled PCR products were bounded to streptoavidin-coated wells of a microtiter plate and detected by anti-DIG–peroxidase conjugate. The biotin-labeled internal probe was used for verification of PCR products. Results: Heat-labile toxin was detected by PCR-ELISA method. The sensitivity of heat-labile toxin was 1.9 ng. This method did not cross-react with bacteria from this variety. Conclusion: PCR-ELISA method is 100 times more sensitive than conventional PCR method and due to lack of agarose gel and electrophoresis device it can be a good alternative to traditional method