ABSTRACT
Three new structurally related depsipeptides, halicylindramides F-H (1-3), and two known halicylindramides were isolated from a Petrosia sp. marine sponge collected off the shore of Youngdeok-Gun, East Sea, Republic of Korea. Their planar structures were elucidated by extensive spectroscopic data analyses including 1D and 2D NMR data as well as MS data. The absolute configurations of halicylindramides F-H (1-3) were determined by Marfey's method in combination with Edman degradation. The absolute configurations at C-4 of the dioxyindolyl alanine (Dioia) residues of halicylindramides G (2) and H (3) were determined as 4S and 4R, respectively, based on ECD spectroscopy. The C-2 configurations of Dioia in 2 and 3 were speculated to both be 2R based on the shared biogenesis of the halicylindramides. Halicylindramides F (1), A (4), and C (5) showed human farnesoid X receptor (hFXR) antagonistic activities, but did not bind directly to hFXR.
Subject(s)
Depsipeptides , Petrosia/chemistry , Receptors, Cytoplasmic and Nuclear/drug effects , Animals , Depsipeptides/chemistry , Depsipeptides/isolation & purification , Depsipeptides/pharmacology , Humans , Marine Biology , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Republic of KoreaABSTRACT
BACKGROUND: Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is a powerful method for the analysis of gene expression. Target gene expression levels are usually normalized to a consistently expressed reference gene also known as internal standard, in the same sample. However, much effort has not been expended thus far in the search for reference genes suitable for the study of stomach cancer using RT-qPCR, although selection of optimal reference genes is critical for interpretation of results. METHODS: We assessed the suitability of six possible reference genes, beta-actin (ACTB), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyl transferase 1 (HPRT1), beta-2-microglobulin (B2M), ribosomal subunit L29 (RPL29) and 18S ribosomal RNA (18S rRNA) in 20 normal and tumor stomach tissue pairs of stomach cancer patients and 6 stomach cancer cell lines, by RT-qPCR. Employing expression stability analyses using NormFinder and geNorm algorithms we determined the order of performance of these reference genes and their variation values. RESULTS: This RT-qPCR study showed that there are statistically significant (p < 0.05) differences in the expression levels of HPRT1 and 18S rRNA in 'normal-' versus 'tumor stomach tissues'. The stability analyses by geNorm suggest B2M-GAPDH, as best reference gene combination for 'stomach cancer cell lines'; RPL29-HPRT1, for 'all stomach tissues'; and ACTB-18S rRNA, for 'all stomach cell lines and tissues'. NormFinder also identified B2M as the best reference gene for 'stomach cancer cell lines', RPL29-B2M for 'all stomach tissues', and 18S rRNA-ACTB for 'all stomach cell lines and tissues'. The comparisons of normalized expression of the target gene, GPNMB, showed different interpretation of target gene expression depend on best single reference gene or combination. CONCLUSION: This study validated RPL29 and RPL29-B2M as the best single reference genes and combination, for RT-qPCR analysis of 'all stomach tissues', and B2M and B2M-GAPDH as the best single reference gene and combination, for 'stomach cancer cell lines'. Use of these validated reference genes should provide more exact interpretation of differential gene expressions at transcription level in stomach cancer.
Subject(s)
Blood Coagulation Factors/genetics , Gene Expression Profiling , Genetic Markers , Glyceraldehyde-3-Phosphate Dehydrogenases/genetics , Reverse Transcriptase Polymerase Chain Reaction , Stomach Neoplasms/genetics , beta 2-Microglobulin/genetics , Algorithms , Cell Line, Tumor , Gene Expression Profiling/standards , Gene Expression Regulation, Neoplastic , Humans , RNA Stability , RNA-Binding Proteins , Reference Standards , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction/standards , Ribosomal ProteinsABSTRACT
Seven new beta-carboline-based metabolites, designated as eudistomins Y1-Y7 ( 1- 7), were isolated from a tunicate of the genus Eudistoma collected near Tong-Yeong City, South Sea, Korea. These new metabolites differ from previously isolated marine metabolites due to the presence of a benzoyl group attached to the beta-carboline nucleus at C-1. Eudistomins Y 1-Y 7 were evaluated for their antibacterial activity, and eudistomin Y6 exhibited moderate antibacterial activity against Gram-positive bacteria Staphylococcus epidermis and Bacillus subtilis without cytotoxicity in the MTT assay at 100 microM.
Subject(s)
Alkaloids/isolation & purification , Anti-Bacterial Agents/isolation & purification , Carbolines/isolation & purification , Gram-Positive Bacteria/drug effects , Urochordata/chemistry , Alkaloids/chemistry , Alkaloids/pharmacology , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Bacillus subtilis/drug effects , Carbolines/chemistry , Carbolines/pharmacology , Korea , Microbial Sensitivity Tests , Molecular Structure , Staphylococcus epidermidis/drug effectsABSTRACT
Three new scalarane-based sesterterpenes, 1- 3, were isolated from a marine sponge of the genus Spongia, and their chemical structures were elucidated by analysis of HRMS and 2-D NMR spectra. The isolated compounds 1 and 3 showed inhibition against the farnesoid X-activated receptor (FXR) with IC50 values of 2.4 and 24 microM, respectively. In particular, compound 3 directly inhibited the interaction between FXR and a coactivator peptide (SRC-1) as determined by surface plasmon resonance (SPR) spectroscopy.
