40 research outputs found
The effect of Chaetocin on the transcriptions of AcMNPV immediate early genes <i>IE0/1</i> (A) and <i>IE2</i> (B).
<p>The error bars indicate the standard deviation calculated from at least three independent parallel experiments. p.i.: post infection.</p
Schematic diagram of <i>Spodoptera</i> Su(<i>var</i>) <i>3-9/eIF2γ</i> genome context and splice pattern.
<p>Numbered boxes represent exons of ORFs. The corresponding sites in genome for each exon are indicated (numbers at upper left for starts and lower right for ends). Broken lines between exons represent intron regions. The polyadenylation signals downstream of ORFs are indicated.</p
Activity of Su(var) 3-9 in Sf9 cells.
<p>Sf9 cells were harvested 3 d post treatments with Chaetocin at appropriate concentration determined by MTT assay (A) and transient overexpression of S. <i>frugiperda</i> Su(var) 3-9. Nuclear proteins were extracted and the global methylation level was evaluated (B). Cellular DNA was isolated and digested with DNase I. The percentage of DNA resistant to the nuclease was determined with qPCR with primers for S. <i>frugiperda β-Actin</i> and β<i>-Tubulin</i> (C, D). The error bars indicate the standard deviation calculated from at least three independent parallel experiments.</p
DataSheet_3_Expression and significance of m6A-RNA-methylation in oral cancer and precancerous lesion.pdf
BackgroundOral potentially malignant disorders (OPMDs) and oral squamous cell carcinoma (OSCC) are a series of related pathologic and molecular events involving simple epithelial hyperplasia, mild to severe dysplasia and canceration. N6-methyladenosine RNA methylation, as the most common modification of both coding mRNA and non-coding ncRNA in eukaryotes, participates in the regulation of the occurrence and development of various malignant tumors in human. However, its role in oral epithelial dysplasia (OED) and OSCC remain unclear.Materials and methodsIn this study, multiple public databases were used for bioinformatics analysis of 23 common m6A methylation regulators in head and neck squamous cell carcinoma (HNSCC). Protein expressions of IGF2BP2 and IGF2BP3 were verified accordingly in clinical cohort samples of OED and OSCC.ResultsPatients with high expression of FTO、HNRNPC、HNRNPA2B1、LRPPRC、IGF2BP1、IGF2BP2、IGF2BP3 had a poor prognosis. IGF2BP2 had a relatively high mutation rate in HNSCC, and its expression was significantly positively correlated with tumor purity, and significantly negatively correlated with the infiltration level of B cells and CD8+T cells. The expression of IGF2BP3 was significantly positively correlated with tumor purity and CD4+T cells. Immunohistochemistrically, the expression of IGF2BP2 and IGF2BP3 in oral simple epithelial hyperplasia, OED and OSCC increased gradually. Both were strongly expressed in OSCC.ConclusionIGF2BP2 and IGF2BP3 were the potential biological prognostic indicators of OED and OSCC.</p
DataSheet_1_Expression and significance of m6A-RNA-methylation in oral cancer and precancerous lesion.pdf
BackgroundOral potentially malignant disorders (OPMDs) and oral squamous cell carcinoma (OSCC) are a series of related pathologic and molecular events involving simple epithelial hyperplasia, mild to severe dysplasia and canceration. N6-methyladenosine RNA methylation, as the most common modification of both coding mRNA and non-coding ncRNA in eukaryotes, participates in the regulation of the occurrence and development of various malignant tumors in human. However, its role in oral epithelial dysplasia (OED) and OSCC remain unclear.Materials and methodsIn this study, multiple public databases were used for bioinformatics analysis of 23 common m6A methylation regulators in head and neck squamous cell carcinoma (HNSCC). Protein expressions of IGF2BP2 and IGF2BP3 were verified accordingly in clinical cohort samples of OED and OSCC.ResultsPatients with high expression of FTO、HNRNPC、HNRNPA2B1、LRPPRC、IGF2BP1、IGF2BP2、IGF2BP3 had a poor prognosis. IGF2BP2 had a relatively high mutation rate in HNSCC, and its expression was significantly positively correlated with tumor purity, and significantly negatively correlated with the infiltration level of B cells and CD8+T cells. The expression of IGF2BP3 was significantly positively correlated with tumor purity and CD4+T cells. Immunohistochemistrically, the expression of IGF2BP2 and IGF2BP3 in oral simple epithelial hyperplasia, OED and OSCC increased gradually. Both were strongly expressed in OSCC.ConclusionIGF2BP2 and IGF2BP3 were the potential biological prognostic indicators of OED and OSCC.</p
Identification and Characterization of the <i>Spodoptera</i> Su(var) 3-9 Histone H3K9 trimethyltransferase and Its Effect in AcMNPV Infection
<div><p>Histone H3-lysine<sup>9</sup> (H3K9) trimethyltransferase gene <i>Su</i>(<i>var</i>) <i>3-9</i> was cloned and identified in three <i>Spodoptera</i> insects, <i>Spodoptera</i><i>frugiperda</i> (<i>S</i><i>. frugiperda</i>), <i>S</i><i>. exigua</i> and <i>S</i><i>. litura</i>. Sequence analysis showed that <i>Spodoptera</i> Su(<i>var</i>) <i>3-9</i> is highly conserved evolutionarily. Su(var) 3-9 protein was found to be localized in the nucleus in Sf9 cells, and interact with histone H3, and the heterochromatin protein 1a (HP1a) and HP1b. A dose-dependent enzymatic activity was found at both 27 °C and 37 °C in vitro, with higher activity at 27 °C. Addition of specific inhibitor chaetocin resulted in decreased histone methylation level and host chromatin relaxation. In contrast, overexpression of Su(var) 3-9 caused increased histone methylation level and cellular genome compaction. In AcMNV-infected Sf9 cells, the transcription of <i>Su</i>(<i>var</i>) <i>3-9</i> increased at late time of infection, although the mRNA levels of most cellular genes decreased. Pre-treatment of Sf9 cells with chaetocin speeded up viral DNA replication, and increased the transcription level of a variety of virus genes, whereas in Sf9 cells pre-transformed with Su(var) 3-9 expression vector, viral DNA replication slow down slightly. These findings suggest that Su(var) 3-9 might participate in the viral genes expression an genome replication repression during AcMNPV infection. It provided a new insight for the understanding virus–host interaction mechanism.</p> </div
Influence of AcMNPV infection on the transcription of several host genes.
<p>Sf9 cells inoculated with AcMNPV at MOI=20 pfu/cell or mock infected were harvested at different time point post infection (p.i.). Total RNA was extracted, reverse transcribed and determined by qPCR. The transcription level of <i>β-Actin</i>, β<i>-Tubulin</i>, <i>Su</i>(<i>var</i>) <i>3-9</i>, <i>HP1a</i> and <i>HP1b</i> were normalized to <i>GAPDH</i>. The error bars indicate the standard deviation calculated from at least three independent infections.</p
Phylogenetic tree of selected HMTs.
<p>Method: Neighbor-Joining, phylogeny test: Bootstrap (1 000 reps), substitution model: amino acid with Jones-Taylor-Thornton (JTT) model. Bootstrap values over 50% are indicated on the top of each node. The orders of insect species are indicated. <i>Spodoptera</i> Su(var) 3-9 is highlighted with prism. Abbreviations: Sf, <i>Spodoptera</i><i>frugiperda</i>; Sl<i>, </i><i>Scoliopteryx</i><i>libatrix</i>; Bm, <i>Bombyx mori</i>; Tc, <i>Tribolium</i><i>castaneum</i>; Am, <i>Apis mellifera</i>; Dm, <i>Drosophila melanogaster</i>; Hs, <i>Homo sapiens</i>.</p
The effect of Chaetocin (A) and overexpression of Su(var) 3-9 (B) on AcMNPV DNA replication.
<p>The error bars indicate the standard deviation calculated from at least three independent parallel experiments. The asterisks indicate statistical significances between groups evaluated with student’s <i>t</i> test (∗P < 0.05; ∗∗P < 0.01).</p
DataSheet_2_Expression and significance of m6A-RNA-methylation in oral cancer and precancerous lesion.pdf
BackgroundOral potentially malignant disorders (OPMDs) and oral squamous cell carcinoma (OSCC) are a series of related pathologic and molecular events involving simple epithelial hyperplasia, mild to severe dysplasia and canceration. N6-methyladenosine RNA methylation, as the most common modification of both coding mRNA and non-coding ncRNA in eukaryotes, participates in the regulation of the occurrence and development of various malignant tumors in human. However, its role in oral epithelial dysplasia (OED) and OSCC remain unclear.Materials and methodsIn this study, multiple public databases were used for bioinformatics analysis of 23 common m6A methylation regulators in head and neck squamous cell carcinoma (HNSCC). Protein expressions of IGF2BP2 and IGF2BP3 were verified accordingly in clinical cohort samples of OED and OSCC.ResultsPatients with high expression of FTO、HNRNPC、HNRNPA2B1、LRPPRC、IGF2BP1、IGF2BP2、IGF2BP3 had a poor prognosis. IGF2BP2 had a relatively high mutation rate in HNSCC, and its expression was significantly positively correlated with tumor purity, and significantly negatively correlated with the infiltration level of B cells and CD8+T cells. The expression of IGF2BP3 was significantly positively correlated with tumor purity and CD4+T cells. Immunohistochemistrically, the expression of IGF2BP2 and IGF2BP3 in oral simple epithelial hyperplasia, OED and OSCC increased gradually. Both were strongly expressed in OSCC.ConclusionIGF2BP2 and IGF2BP3 were the potential biological prognostic indicators of OED and OSCC.</p