ABSTRACT
Burkitt lymphoma (BL) is a highly malignant non-Hodgkin's lymphoma that is closely related to the abnormal expression of genes. Familial acute myelogenous leukemia related factor (FAMLF; GenBank accession No. EF413001.1) is a novel gene that was cloned by our research group, and miR-181b is located in the intron of the FAMLF gene. To verify the role of miR-181b and FAMLF in BL, RNAhybrid software was used to predict target site of miR-181b on FAMLF and real-time quantitative PCR (RQ-PCR) was used to detect expression of miR-181b and FAMLF in BL patients, Raji cells and unaffected individuals. miR-181b was then transfected into Raji and CA46 cell lines and FAMLF expression was examined by RQ-PCR and western blotting. Further, Raji cells viability and proliferation were detected by MTT and clone formation, and Raji cell cycle and apoptosis were detected by flow cytometry. The results showed that miR-181b can bind to bases 21-42 of the FAMLF 5' untranslated region (UTR), FAMLF was highly expressed and miR-181b was lowly expressed in BL patients compared with unaffected individuals. FAMLF expression was significantly and inversely correlated to miR-181b expression, and miR-181b negatively regulated FAMLF at posttranscriptional and translational levels. A dual-luciferase reporter gene assay identified that the 5' UTR of FAMLF mRNA contained putative binding sites for miR-181b. Down-regulation of FAMLF by miR-181b arrested cell cycle, inhibited cell viability and proliferation in a BL cell line model. Our findings explain a new mechanism of BL pathogenesis and may also have implications in the therapy of FAMLF-overexpressing BL.
Subject(s)
Burkitt Lymphoma/genetics , Burkitt Lymphoma/metabolism , Gene Expression Regulation, Neoplastic , MicroRNAs/metabolism , Proteins/metabolism , Adolescent , Adult , Apoptosis/genetics , Cell Line, Tumor , Cell Proliferation/genetics , Cell Survival/genetics , Child , Child, Preschool , Down-Regulation/genetics , Female , Humans , Infant , Male , MicroRNAs/genetics , Proteins/genetics , Young AdultABSTRACT
Tea (Camellia sinensis L.) is a thermophilic evergreen woody plant that has poor cold tolerance. The SAD gene plays a key role in regulating fatty acid synthesis and membrane lipid fluidity in response to temperature change. In this study, full-length SAD cDNA was cloned from tea leaves using rapid amplification of cDNA ends and polymerase chain reaction (PCR)-based methods. Sequence analysis demonstrated that CsSAD had a high similarity to other corresponding cDNAs. At 25°C, the CsSAD transcriptional level was highest in the leaf and lowest in the stem, but there was no obvious difference between the root and stem organs. CsSAD expression was investigated by reverse transcription-PCR, which showed that CsSAD was upregulated at 4° and -5°C. At 25°C, CsSAD was induced by polyethylene glycol, abscisic acid, and wounding, and a similar trend was observed at 4°C, but the mean expression level at 4°C was lower than that at 25°C. Under natural cold acclimation, the 'CsCr05' variety's CsSAD expression level increased before decreasing. The CsSAD expression level in variety 'CsCr06' showed no obvious change at first, but rapidly increased to a maximum when the temperature was very low. Our study demonstrates that CsSAD is upregulated in response to different abiotic conditions, and that it is important to study the stress resistance of the tea plant, particularly in response to low temperature, drought, and wounding.
Subject(s)
Adaptation, Physiological , Camellia sinensis/enzymology , Gene Expression Regulation, Plant , Plant Proteins/genetics , Stearoyl-CoA Desaturase/genetics , Amino Acid Sequence , Camellia sinensis/genetics , Camellia sinensis/physiology , Cloning, Molecular , Cold Temperature , Droughts , Molecular Sequence Data , Phylogeny , Plant Components, Aerial/enzymology , Plant Components, Aerial/physiology , Plant Proteins/chemistry , Plant Proteins/metabolism , Plant Roots/enzymology , Plant Roots/physiology , Sequence Alignment , Stearoyl-CoA Desaturase/chemistry , Stearoyl-CoA Desaturase/metabolismABSTRACT
The spermidine synthase (SPDS) gene exists widely in all types of plants. In this paper, the codon usage of the SPDS gene from Camellia sinensis (CsSPDS) was analyzed. The results showed that the codon usage of the CsSPDS gene is biased towards the T-ended or A-ended codons, which is similar to that observed in 73 genes selected from the C. sinensis genome. An ENC-plot for 15 SPDS genes from various plant species suggested that mutational bias was the major factor in shaping codon usage in these genes. Codon usage frequency analysis indicated that there was little difference between the CsSPDS gene and dicot genomes, such as Arabidopsis thaliana and Nicotiana tabacum, but significant differences in codon usage were observed between the CsSPDS gene and monocot genomes, such as Triticum aestivum and Zea mays. Therefore, A. thaliana and N. tabacum expression systems may be more suitable for the expression of the CsSPDS gene.
Subject(s)
Camellia sinensis/enzymology , Camellia sinensis/genetics , Codon , Spermidine Synthase/genetics , Cluster Analysis , Gene Expression Regulation, Plant , Genome, PlantABSTRACT
Numerous studies have evaluated the association between the human interleukin-10 gene -592C>A polymorphism and gastric cancer risk. However, the results have been inconsistent. This meta-analysis was designed to resolve these controversies. Systematic searches of the electronic databases Embase, PubMed, and Google Scholar were performed to identify relevant studies. A meta-analysis was performed to examine the association between the interleukin-10 gene -592C>A polymorphism and gastric cancer risk. Odds ratios (OR) and its 95% confidence intervals (CI) were used for statistical analysis. Twelve studies were included in the meta-analysis, which included 2116 gastric cancer cases and 4077 controls. No significant association was found between the interleukin-10 gene -592C>A polymorphism and gastric cancer risk in total population analysis. In stratified analysis, a significant association was found in the Asian subgroup (AA vs AC: OR = 0.79, 95%CI = 0.64-0.98; dominant model: OR = 1.26, 95%CI = 1.04-1.57), and no significant association was observed among Caucasians. In addition, the corresponding pooled ORs were not substantially altered after excluding one study that deviated from Hardy-Weinberg equilibrium in the control group. This meta-analysis supports an association between the interleukin-10 gene -592C>A polymorphism and gastric cancer risk in Asians but not in Caucasians. Further large and well-designed studies are needed to confirm these conclusions.