Comparison of the expression of human equilibrative nucleotide transporter 1 (hENT1) and ribonucleotide reductase subunit M1 (RRM1) genes in seven non-Hodgkin lymphoma cell lines.

We investigated the variability in the expression of human equilibrative nucleoside transporter 1 (hENT1) and ribonucleotide reductase subunit M1 (RRM1) in non-Hodgkin lymphoma cell lines. hENT1 and RRM1 mRNA expression levels in natural killer (NK) cells and seven non-Hodgkin lymphoma cell lines (YTS, SNK-6, Jeko-1, ly-1, Raji, Karpas, and Jurket) were studied using reverse-transcription quantitative real-time polymerase chain reaction (RT-qPCR) and the results were compared using the Student t-test. mRNA expression of hENT1 was detectable in YTS, SNK-6, Jeko-1, ly-1, Raji, Karpas, Jurket, and NK cells, which revealed variability in gene expression. There were significant differences in the mRNA expression values of hENT1 (P = 0.021) and RRM1 (P = 0.002) compared to those in NK cells. mRNA expression of both hENT1 and RRM1 was closely associated with non-Hodgkin lymphoma cell proliferation. Differential expression analysis of hENT1 and RRM1 in non-Hodgkin lymphoma cell lines may provide novel drug leads for precision medicine.

Effects of cyclopamine on the biological characteristics of human breast cancer MCF-7 cell line and its mechanism.

PURPOSE: To observe the effects of cyclopamine on the biological characteristics of human breast cancer MCF-7 cell line and explore its mechanism. MATERIALS AND METHODS: After human breast cancer MCF-7 cells were treated with different-concentration cyclopamine for different periods, MTT assay was used to detect the inhibitory effect of cyclopamine on MCF-7 cell proliferation, flow cytometry was used to determine the distribution of MCF-7 cell cycle and the effect of cyclopamine on MCF-7 apoptosis, and Western blot was used to measure the protein levels of cyclins D1 and p21 in MCF-7 cells. RESULTS: In certain range, MCF-7 cell proliferation was inhibited by cyclopamine in a dose- and time-dependent manner, and the optimal inhibiting concentration was ten µmol/L and the optimal action time at 48 hours. With the time prolongation of cyclopamine action, the cells in G0/G1 phase were significantly increased, but the cells in S phase were significantly decreased (compared with blank control group, allp < 0.05). With the time prolongation of cyclopamine action, apoptosis rate of MCF-7 cells was also significantly increased (compared with blank control group, allp < 0.05). The level of cyclin D1 of MCF-7 cells was decreased, but cyclin p21 was increased (compared with blank control group, all p < 0.05). CONCLUSION: Cyclopamine inhibits MCF-7 cell proliferation via arresting MCF-7 cell transformation from G1 phase to S phase. This may be associated with the expressions of Hedgehog (Hh) signaling pathway-related cyclins.

Selenium supplementation of children in a selenium-deficient area in China: blood selenium levels and glutathione peroxidase activities.

Keshan disease is a cardiomyopathy restricted to the endemic areas of China and seen in residents having an extremely low selenium (Se) status. Prophylactic administration of sodium selenite has been shown to decrease significantly the incidence of acute and subacute cases. The aim offthe study was to assess the relative bioavailability of selenite versus organic Se-yeast in a Se-deficient area in China with a randomized double-blind double-dummy design. Healthy children (n=30) between 14 and 16 yr of age were randomized into three equal groups receiving either 200 microg/d selenite Se or 200 microg/d Se-yeast or placebo for 12 wk. Blood was drawn at baseline, 4, 8, and 12 wk and 4 wk postsupplementation. The plasma Se concentration (mean +/- SD) was 0.16+/-0.03 micromol/L at baseline. Selenite and Se-yeast supplementation increased plasma Se to plateau values, 1.0+/-0.2 and 1.3+/-0.2 micromol/L, respectively. In red cells, Se-yeast increased the selenium level sixfold and selenite threefold compared to placebo. The relative bioavailability of Se-yeast versus selenite measured as glutathione peroxidase (GSHPx) activity was similar in plasma, red blood cells, and platelets. GSHPx activity reached maximal levels in plasma and platelets of 300% and 200%, respectively, after 8 wk compared to the placebo group, but continued to increase in red cells for 16 wk. Our study showed that although both forms of Se were equally effective in raising GSHPx activity, Se-yeast provided a longer lasting body pool of Se. Se-yeast may be a better alternative to selenite in the prophylaxis of Keshan disease with respect to building up of body stores.

Further investigation on the role of selenium deficiency in the aetiology and pathogenesis of Keshan disease.

Selenium supplements were not able to restore the ultrastructural changes in the myocardium of latent Keshan disease patients taken by using cardiac catheter endomyocardial biopsy. Observations on the changes of selenium status and the incidence of Keshan disease showed that new latent and naturally-occurring chronic cases were found in the endemic area even after selenium levels had been elevated in the residents to the levels typical in the non-endemic area. These results indicate that although selenium deficiency might be a primary pathogenetic geogen in the occurrence of Keshan disease, it is rather a conditional predisposing factor than a specific or initiative aetiologic factor for the occurrence of Keshan disease. Selenium supplementation could apparently alleviate the higher platelet responsiveness of residents in the endemic area, which might contribute to eliminating the basis for the occurrence of the multifocal perivascular necroses in myocardium of acute and subacute Keshan disease.

Synthesis and characterization of monohydroxylated derivatives of 7H-dibenzo[c,g]carbazole.

The synthesis of several monohydroxylated derivatives of the potent carcinogen 7H-dibenzo[c,g]carbazole (DBC), including 1-hydroxy-7H-dibenzo[c,g]carbazole (1-OH-DBC), 13-c-hydroxydibenzo[c,g]carbazole (13-c-OH-DBC), and 5-hydroxy-7H-dibenzo[c,g]carbazole (5-OH-DBC), is described. 1-OH-DBC was prepared from 8-methoxy-2-tetralone and 2-naphthyl-hydrazine via Fischer indole synthesis followed by boron tribromide demethylation. The rearrangement and hydrolysis reactions to give 13-c-OH-DBC from DBC and benzoyl peroxide are discussed. The preparation and isolation of 5-OH-DBC, by hydrolysis of 5-acetoxy-N-acetyl-DBC, and the formation of its intermediate 5-acetoxy-DBC and its byproduct 6,6'-bis-(5-OH-DBC) are described in detail.

Comparative metabolism of 7H-dibenzo[c,g]carbazole and dibenz[a,j]acridine by mouse and rat liver microsomes.

The comparative metabolism of the carcinogenic pollutants 7H-dibenzo[c,g]-carbazole (DBC) and dibenz[a,j]acridine (DBA) was investigated in vitro using 3-methylcholanthrene (3MC) induced Sprague-Dawley rat and Hsd:ICR(Br) mouse liver microsomal preparations with benzo[a]pyrene (BaP) as the positive control. Metabolites were isolated and separated by HPLC and identified by spectroscopic and co-chromatographic techniques using synthetic standards. The major metabolites of DBC were the phenols: the 5-OH-DBC, 3-OH-DBC, and 2-OH-DBC. Traces of 1-OH-DBC were also found yet no dihydrodiols were identified. The major metabolites of DBA were the 3,4-diol-DBA and 5,6-diol-DBA, 1,2-diol-DBA, DBA-5,6-oxide and 4-OH-DBA. Treatment of both mice and rats with 3MC resulted in significant (P less than or equal to 0.05) increases relative to control in the microsomal metabolism of DBA to dihydrodiol and phenol metabolites, similar to that observed for BaP. 3MC-induced rat liver microsomes significantly (P less than or equal to 0.05) increased DBC metabolism relative to control microsomes whereas DBC metabolism was not increased with 3MC-induced mouse liver microsomes. These data indicate that different enzymatic pathways are involved in the metabolic activation of DBC in the Hsd:ICR(Br) mouse and Sprague-Dawley rat.

The identification and quantitation of alkylated nucleobases by high-performance liquid chromatography with UV photodiode array detection.

The application of UV diode array detection in high-performance liquid chromatographic (HPLC) identification and quantitation of several classes of synthetic and commercially available alkylated nucleobases is investigated. Quantitative spectral overlays of these compounds to methyl standard references from a spectral library and absorbance ratios at two maximal wavelengths (lambda max) are found to be useful in categorizing the solutes. They can be grouped into classes of compounds originating from a specific nucleobase and classes of analogs having different alkyl substituents (e.g., methyl, ethyl, propyl, allyl, and benzyl) at the same position of the heterocycle. At a selected wavelength for alkylated nucleobases in the same class, the detector response factors are independent of the alkyl group (+/- 10%). This technique provides a practical means for both qualitative and quantitative analysis of product distribution of DNA base alkylation by using only readily obtainable methylated derivatives as the reference standards.

Separation characteristics of alkylated guanines in high-performance liquid chromatography.

The retention behavior of thirteen alkylated guanines on normal-phase silica gel and amino columns and on reversed-phase ODS and phenyl columns was studied. The larger the alkyl substituent at the same position of guanine the weaker was the retention in the normal-phase chromatographic system and the greater the retention during reversed-phase chromatography. O6-Derivatives possess the lowest polarity in each set of isomers. An amino column was found to be of highest efficiency in terms of separation of the set of ethylguanine isomers and of benzylguanines studied. A phenyl column provided the best resolution of methylated guanines.