Detection of free DNA septin 9 gene methylation in plasma / 中南大学学报(医学版)

OBJECTIVES@#To explore the correlation between cytosine-phosphoric-guanylic (CpG) site of Septin 9 gene and colorectal cancer, and to develop a real-time PCR detection system in plasma in patients with colorectal cancer.@*METHODS@#The methylation of training samples was detected by high-throughput sequencing technology, and the sites highly consistent with the clinical information of colorectal cancer were identified. Then the detection system of real-time PCR was designed to analyze the consistency of plasma and tissue based on methylationa sensitive enzyme digestion. Finally, 100 clinical trials were conducted to evaluate the performance of the detection system with the methylation sensitive enzyme digestion-real-time PCR.@*RESULTS@#The highly consistent sites, which were selected by high-throughput sequencing from 71 training set samples, was the 38th CpG. Based on the detection region, the screened methylation sensitive enzymes were @*CONCLUSIONS@#The 38th CpG site of Septin 9 detected by the detection system of methylation sensitive enzyme digestion-real-time PCR can highly predict the occurrence of colorectal cancer with great clinical application value.

MLL-SEPT5 Fusion Transcript in Two de novo Acute Myeloid Leukemia Patients With t(11;22)(q23;q11)

No abstract available.

Septin 7: Actin cross-organization is required for axonal association of Schwann cells

Myelin sheaths present two distinct domains: compacted myelin spirals and flanking non-compacted cytoplasmic channels, where lipid and protein segregation is established by unknown mechanisms. Septins, a conserved family of membrane and cytoskeletal interacting GTPases, form intracellular diffusion barriers during cell division and neurite extension and are expressed in myelinating cells. Septins, particularly septin 7 (Sept7), the central constituent of septin polymers, are associated with the cytoplasmic channels of myelinating cells. Here we show that Schwann cells deprived of Sept7 fail to wrap around axons from dorsal root ganglion neurons and exhibit disorganization of the actin cytoskeleton. Likewise, Sept7 distribution is dependent on microfilament but not microtubule organization.

Preliminary analysis of differentially expressed proteins of clear-cell renal cell carcinoma by comparative proteomic technologies / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the different expression of proteins between human clear-cell renal cell carcinoma (ccRCC) cell line RLC-310 and normal renal proximal tubule epithelial cell line HK-2, and to search new differentially expressed proteins of RCC.</p><p><b>METHODS</b>RLC-310 and HK-2 cells were cultured in vitro. The total proteins were separated by ProteomeLab PF 2D protein fractionation system and the differential expression protein fractions of the two cell lines were analyzed and identified by capillary liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS). RT-PCR and Western blot were used to confirm the representative differential expression at mRNA and protein levels.</p><p><b>RESULTS</b>One hundred and ninty-six differentially expressed proteins were identified. These differentially expressed proteins involved in many aspects, including cell proliferation and anti-apoptosis, energy metabolism, mitochondria reduction and oxidation, oxidative stress and resistance, cell signaling, invasion and adhesion, cytoskeleton and motion, neovascularization, etc. Except for previously reported RCC associated proteins: annexin A2, fatty acid-binding protein, vimentin, fibronectin, and so on, Septin-9 was firstly found highly expressed in RLC-310 cells when compared with that in the HK-2 cells. Moreover, the overexpression of Septin-9 was confirmed by RT-PCR and Western blot analysis at both mRNA and protein levels (P < 0.05).</p><p><b>CONCLUSIONS</b>The human ccRCC cell line RLC-310 cells display differential protein profiles compared with those of the normal renal cell line HK-2 cells. The identified differential expression proteins are involved in many aspects of RCC development. It is worth further study and elucidate the molecular mechanisms of RCC. The representative differential protein Septin-9 deserves further study its role in the angiogenesis of ccRCC.</p>

Expression of SEPT4 protein in the ejaculated sperm of idiopathic asthenozoospermic men / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the role of the SEPT4 protein in the pathogenesis of idiopathic asthenozoospermia.</p><p><b>METHODS</b>Samples of ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men were separated and purified by Percoll discontinuous density gradients, the distribution and expression of SEPT4 in the sperm samples were determined by immunocytochemistry, and the expressions of SEPT4 mRNA and SEPT4 protein were detected by RT-PCR and Western blot.</p><p><b>RESULTS</b>Immunocytochemistry showed that the expression of SEPT4, located in the annulus, was significantly reduced in the sperm of the idiopathic asthenozoospermia patients (t = 3.452, P < 0.01). RT-PCR revealed that the expression of SEPT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (t = 3.521, P < 0.05). Western blot confirmed the results of RT-PCR (t = 5.872, P < 0.05).</p><p><b>CONCLUSION</b>The expression of SEPT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.</p>

Update of asthenospermia-related genes and proteins / 中华男科学杂志

One of the most common causes of male infertility is asthenospermia, whose pathogenesis, however, is not yet clear. Recent researches have found that some genes (such as tektin-2, DNAI1, DNAH5, DNAH11, AKAP4, SEPT4 and Smcp) and proteins (such as sperm proteins ACTB, ANXA5, PRM1, PRM2 and SABP and seminal proteins Tf, PSA, PAP and Fractalkine) are associated with asthenospermia. The finding of these molecular markers has provided a base for the explanation of the molecular mechanism of asthenospermia, and these markers may become the diagnostic and therapeutic targets of the disease.

Study on the anti-invasion effect of SEPT7 gene for U251MG glioma cell in vitro / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the anti-invasion effect of SEPT7 gene on U251MG glioma cells and its possible molecular mechanism.</p><p><b>METHODS</b>Recombinant adenovirus vector carrying SEPT7 gene (rAd5-SEPT7) was transduced to human glioma cell line U251MG, and empty adenovirus vector was used as control. Tumor invasion was examined by Transwell method and 3 D-Matrigel assay, and tumor cell migration by wound-healing method and 2 D-Matrigel assay. Three major molecular events associated with cell motility and migration, including changes of expression in MMP2, MMP9, MT1-MMP, TIMP1 and TIMP2, the alteration of integrin alpha(v)beta(3) expression, and the structural change of cytoskeleton protein, tubulin-alpha, in U251 cells transduced with rAd5-SEPT7 were studied by Western blotting, immunofluorescence and laser scanning confocal microscope, respectively.</p><p><b>RESULTS</b>The invasive and migratory capabilities of cells transduced with rAd5-SEPT7 were inhibited. The expression of extracellular matrix metalloproteinases MMP-2, MMP-9, MT1-MMP and integrin alpha(v)beta(3) was significantly decreased, while the expression of matrix metalloproteinase inhibitor TIMP1, TIMP2 was upregulated. Intracellular cytoskeleton protein-tubulin-alpha in U251 cells exhibited prominent morphological changes which including the appearance of distortion and aggregation resulting from redistribution of tubulin-alpha, and this feature of alteration was similar to the tubulin-alpha structure in normal non-tumor cells.</p><p><b>CONCLUSION</b>SEPT7 gene can inhibit the invasion and migration ability of U251 glioma cells. Its molecular mechanism may include that SEPT7 gene reverses the imbalanced state of MMPs/TIMPs, downregulates the expression of integrin alpha(v)beta(3) and alters the structure of tubulin-alpha of U251MG glioma cells. It is suggested that SEPT7 gene could be a good candidate for gene therapy of gliomas.</p>

An observation of the effects of the truncated septin2 on mouse epidermal cell and fibroblast / 中华整形外科杂志

<p><b>OBJECTIVE</b>To construct eukaryotic expression vector of the truncated septin2 and investigate the influence on the cultured mouse epidermal cell and fibroblast in vitro exerted by the transgenic expression product.</p><p><b>METHODS</b>The short splicing fragment was obtained by amplifying the reverse transcription product of the fetal mouse skin mRNA with PCR. Then its recombinant expression vector pcDNA3.1 (-)/septin2s was constructed and used to transfect the mouse epidermal cell and fibroblast cultured in vitro. The expression of the foreign gene was detected with RT-PCR and the changes of cell proliferation were observed and analysed.</p><p><b>RESULTS</b>RT-PCR results indicated that pcDNA3.1/septin2 was expressed in the cultured mouse epidermal cells and fibroblasts in vitro. We found that the epidermal cells accelerated their reproduction, but the fibroblasts had no obvious changes.</p><p><b>CONCLUSION</b>We successfully constructed eukaryotic expressive vectors of pcDNA3.1/ septin2s and transfected it into mouse epidermal cells and fibroblasts in vitro. The results settle a basis for showing effect of septin2s on fetal mouse skin.</p>

Influence of SEPT7 on biological characters of glioma cell line TJ905 / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the influence of SEPT7 on biological characters of gliomas cells TJ905.</p><p><b>METHODS</b>Recombinant SEPT7 constructs was transfected to human glioblastoma cell line TJ905 in which SEPT7 expression is absent. The positive clones were identified by RT-PCR and Western blot analysis. The cell proliferation was determined by MTT assay and flow cytometry, cell apoptosis was detected with Annexin V staining and cell invasion was evaluated by motility in three-dimensional culture. Moreover, the molecules regulating the cell cycle progression were examined by immunofluorescence staining and Western blot analysis.</p><p><b>RESULTS</b>When SEPT7 was successfully transfected to TJ905 cells, the cell proliferation activity of TJ905 cell was inhibited, the cell cycle was arrested in G0/G1 phase and S phase fraction (SPF) was lowered, the positive regulatory molecules for cell cycle progression including cyclin D1, CDk4, cyclin E and CDk2 were downregulated while the negative modulators including p16 and p21 were upregulated, apoptotic cells were increased and cell invasive ability was attenuated.</p><p><b>CONCLUSIONS</b>Transfection of SEPT7 construct into the glioma cells TJ905 is able to inhibit the proliferation activity and invasive ability of TJ905 cell and to induce cell apoptosis. These results revealed that SEPT7 exerted the suppressive effect on the glioma cell growth and invasion, and induced apoptosis, and suggested that SEPT7 as a gene of glioma suppressor.</p>