Analysis and verification of the interaction network of differentially expressed genes in invasive bladder cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the data of gene expression microarray by protein interaction network analysis, establish an interaction network of differentially expressed genes in invasive bladder cancer and verify the central nodes of the network.</p><p><b>METHODS</b>A total of 152 differentially expressed genes in invasive bladder cancer detected by gene expression microarray were inputted into STRING database online for analysis and establishment of the interaction network. The interaction data were imported into Cytoscape 2.6.2 software for screening the central nodes of the network. KEGG database was exploited for pathway analysis and functional study of the central node genes. Real-time RT-PCR was used for verification, and the genes with maximal differential expressions were screened for exploring the molecular mechanism of carcinogenesis of invasive bladder cancer.</p><p><b>RESULTS</b>The protein products of 103 differentially expressed genes in bladder cancer had interactions, forming a complicated interaction network. Twenty-six nodes involved in several signal pathways were confirmed by Cytoscape as the central nodes of the network, among which UBE2C, VEGF, TGFBR2, and CAV1 nodes were verified by real-time RT-PCR as the genes with maximal differential expressions between the bladder cancer and normal tissues, and the 2(-delta delta Ct) of these genes were 9.45, 4.17, 0.13 and 0.18 (GAPHD as the internal control), respectively.</p><p><b>CONCLUSION</b>The interaction network of the differentially expressed genes, especially the central nodes of this network, can provide clues to the carcinogenesis, early diagnosis and molecular targeted therapy of invasive bladder cancer.</p>

Mutation of MTCYB and MTATP6 is associated with asthenospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the correlation of the mutation of MTCYB and MTATP6 genes in sperm mitochondria with asthenospermia.</p><p><b>METHODS</b>We extracted mtDNA from 80 semen samples of asthenospermia and 20 of normal sperm motility, amplified the MTCYB and MTATP6 genes by PCR, and analyzed their mutation by sequencing and BLAST matching.</p><p><b>RESULTS</b>The deletion of both MTCYB and MTATP6 were detected in 20 of the 80 asthenospermia samples, MTCYB deletion in 16 and MTATP6 deletion in 4, accounting for 20% and 5% respectively. Sequencing and BLAST matching revealed G8887A mutation in the MTATP6 gene in the asthenospermia samples, with a mutation rate of 20%, while no regular mutation was noted in MTCYB. Neither significant deletion nor mutation was observed in any of the 20 samples of normal sperm motility.</p><p><b>CONCLUSION</b>Both the deletion and mutation of MTCYB and MTATP6 genes in sperm mitochondria might affect sperm motility in adults.</p>

Effect of vascular endothelial growth factor gene transfer on proliferation and metabolism of human bone marrow stromal cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of vascular endothelial growth factor 165 (VEGF165) gene transfer on the proliferation and metabolism of human bone marrow stromal cells (hBMSCs) in vitro.</p><p><b>METHODS</b>hBMSCs were divided into 3 groups and subjected to adenovirus mediated VEGF165 gene transfection, transfection with empty adenoviral vector, or left untreated (control). MTT assay and flow cytometry were performed to analyze the proliferation of the cells after corresponding treatments. The third passage of hBMSCs (2x10(4)/ml), after corresponding transfection procedures, were cultured in conditional medium and tested for ALP content 2, 4 and 6 days after the transfection. Also at 3, 5 and 7 days after the transfection, the cells were examined for osteocalcin (C) and laminin (LN) contents.</p><p><b>RESULTS</b>The number of cells in each group increased with the culture time without obvious differences in the optical density. No significant differences were noted between the 3 groups in the percentage of G1 phase cells or in the proliferation index (PrI) (P>0.05), but compared with the nontransfected and the empty vector-transfected cells, the cells with VEGF165 gene transfection had significantly higher ALP, OC and LN contents (P<0.05).</p><p><b>CONCLUSION</b>VEGf165 gene transfer does not obviously affect the proliferation of cultured hBMSCs, but can increase the cellular secretion of AIP, C and LN, suggesting that VEGF165 promotes the differentiation of hBMSCs into osteoblasts in vitro.</p>

Screening differentially expressed genes in denucleated K562 cells with restriction display technique / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen differentially expressed genes in cytochalasin B (CB)-induced denucleated K562 cells by restriction display (RD) technique.</p><p><b>METHODS</b>The total RNA was isolated and purified from K562 cells before and after CB (10 mug/ml) treatment. The mRNA from both treated and untreated K562 cells were reversely transcribed into cDNA, and the differentially expressed genes were separated using RD technique combined with polyacrylamide gel electrophoresis and sliver staining, followed by cloning, sequencing and homology analysis against GenBank database of these genes.</p><p><b>RESULTS</b>Seven differentially expressed genes were identified in CB-treated cells including aquaporin 1 (AQP1) gene, which was verified to be up-regulated after CB treatment by RT-PCR.</p><p><b>CONCLUSION</b>AQP1 gene might be in close association with the regulation of denucleation processes and CB-induced proliferation inhibition of K562 cells.</p>

Differential gene expression profile of keloids: a study with cDNA microarray / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differentially expressed genes in keloids in comparison with normal skin using cDNA microarray.</p><p><b>METHODS</b>The cDNA microarray consisting of 8064 clones of human genes was employed to detect and screen the differentially expressed genes in keloid and normal skin tissues. Semi-quantitative RT-PCR was applied to verify the results of gene microarray.</p><p><b>RESULTS</b>Totally 277 differentially expressed genes were identified in keloids in comparison with normal skin tissue, including 163 up-regulated genes and 114 down-regulated ones according to the designed data filter criteria. These differentially expressed genes belonged to 26 different functional gene families involving different biological processes. RT-PCR yielded results were consistent with those of microarray study.</p><p><b>CONCLUSION</b>A variety of genes are involved in the formation of keloids. The 277 differentially expressed genes comprise the differential gene expression profile of keloids and describe the general changes in the gene expressions in keloid at transcriptional level. Further analysis of the identified genes might help reveal the molecular mechanism of abnormal scarring.</p>

Research of the spermatozoal gene expression with gene microarrays / 中华男科学杂志

<p><b>OBJECTIVE</b>To perform the detection of spermatozoal gene expression in order to accelerate the study of spermatozoal molecular biology.</p><p><b>METHODS</b>To collect the healthy adults sperm and lymphocytes respectively, and then to extract the total RNAs from them by RNeasy mini kit (QIAGEN) or Trizol reagent. Corresponding cDNAs were produced, digested, ligated, finally labeled with Cy3 (sperm) and CyS (lymphocyte) in the course of RD amplifying reactions. Hybridization with self-made microarrays contained 560 probes was carried out after the labeled cDNAs pured by PCR Product Purification Kit.</p><p><b>RESULTS</b>Among the 560 probes, 72 genes were up-regulated, 321 genes were down-regulated, the others had no different expression. Furthermore, genes associated with replication, transcription, translation and regulative functions were non-different expression or down-regulated, and those belonged to the spermatogenesis associated, sperm associated antigen were up-regulated, but those involved in the glycolysis were up-regulated, in the oxidative phosphorylation were down-regulated.</p><p><b>CONCLUSION</b>It had successfully confirmed that there were a plenty of genes expressed in sperm, furthermore the genes expressed were accorded to spermatozoal functions and characteristics.</p>