ABSTRACT
Esophageal squamous cell carcinoma (ESCC) is the leading pathologic type in China. miR-145 has been reported to be downregulated in multiple tumors. This study was aimed to investigate the role of miR-145 in ESCC. miR-145 expression was investigated in 65 ESCC samples as well as four ESCC cell lines by quantitative real-time polymerase chain reaction (qRT-PCR). Targetscan 6.2 website (http://www.targetscan.org/) was used to predict the targets of miR-145. Expression of phospholipase C epsilon 1 (PLCE1) messenger RNA and protein was detected by qRT-PCR or Western blot. MTT and wound healing assay were conducted to explore the effects of miR-145 on the proliferation and migration of ESCC cell lines, respectively. miR-145 was significantly decreased in ESCC tissues. An inverse correlation between miR-145 and invasion depth and TNM stage were observed. PLCE1 was a direct target of miR-145, and the expression of PLCE1 was inversely correlated with miR-145 expression in ESCC tissues. In addition, overexpression of miR-145 suppressed cell proliferation and migration in ESCC cells. The enforced expression of PLCE1 partially reversed the suppressive effect of miR-145. These results prove that miR-145 may perform as a tumor suppressor in ESCC by targeting PLCE1.
Subject(s)
Esophageal Neoplasms/genetics , Esophageal Squamous Cell Carcinoma/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Phosphoinositide Phospholipase C/genetics , Aged , Base Sequence , Cell Line, Tumor , Cell Movement , Cell Proliferation , Epithelial Cells/metabolism , Epithelial Cells/pathology , Esophageal Neoplasms/metabolism , Esophageal Neoplasms/pathology , Esophageal Neoplasms/surgery , Esophageal Squamous Cell Carcinoma/metabolism , Esophageal Squamous Cell Carcinoma/pathology , Esophageal Squamous Cell Carcinoma/surgery , Female , Humans , Lymphatic Metastasis , Male , MicroRNAs/metabolism , Middle Aged , Neoplasm Staging , Phosphoinositide Phospholipase C/antagonists & inhibitors , Phosphoinositide Phospholipase C/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Signal TransductionABSTRACT
A novel mono-modified ß-cyclodextrin (ß-CD) consisting of 4-carbamoylmethoxy-benzoyl unit at the primary side was synthesized and its self-assembly behavior was determined by X-ray crystallography and NMR spectroscopy. The crystal structure shows a 'Yin-Yang'-like packing mode, in which the modified ß-CD exhibits a channel superstructure formed by a tail-to-tail dimer as the repeating motif with the substituted group embedded within the hydrophobic cavity of the facing ß-CD. The geometry of the substituted group is determined by the inclusion of the cavity and is further stabilized by two intermolecular hydrogen bonds between the carbonyl O atom and phenyl group. Furthermore, NMR ROESY investigation indicates that the self-assembly behavior of the substituted group within the ß-CD cavity is retained in aqueous solution, and the effective binding constant Ka was calculated to be 1330 M(-1) by means of (1)H NMR titration according to iterative determination.
Subject(s)
Acetamides/chemical synthesis , beta-Cyclodextrins/chemical synthesis , Acetamides/chemistry , Carbohydrate Conformation , Carbohydrate Sequence , Crystallography, X-Ray , Models, Molecular , Molecular Sequence Data , Solutions , beta-Cyclodextrins/chemistryABSTRACT
Environmental stresses, such as salinity, drought and cold, can induce the expression of a large amount of genes. Among these are many transcription factors that regulate the expression of downstream genes by specifically binding to cis-elements or forming transcriptional complexes with other proteins. In the present study, a DREB-like transcription factor gene, named AhDREB1, was isolated from a halophyte Atriplex hortensis. AhDREB1 encoded a protein containing a conserved EREBP/AP2 domain featuring the DREB family. In yeast one-hybrid analysis AhDREB1 protein was specifically bound to DRE elements and activated the expression of the reporter genes of HIS3 and LacZ. The AhDREB1 gene was expressed in roots, stems and leaves of A. hortensis. Salinity induced its expression in roots, but not in other organs. Overexpression of AhDREB1 in transgenic tobacco led to the accumulation of its putative downstream genes. The performance of the transgenic lines was also tested under stressed conditions and two lines were found to be stress-tolerant. These results suggest that the AhDREB1 protein functions as a DRE-binding transcription factor and play roles in the stress-tolerant response of A. hortensis.
