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马端, Bilian Zhao a, Xinping Luo a, *, Haiming Shi a, Duan Ma b, **
Thrombosis Research xxx (2011),-0001,():
-1年11月30日
Introduction: Tissue factor pathway inhibitor-2 (TFPI-2) is a member of the Kunitz-type family of serine protease inhibitors, which inhibits several matrix metalloproteinases activity involved in extracellular matrix degradation. Studies have shown low TFPI-2 expression in the shoulder regions of atherosclerotic plaques. But studies evaluating its role in the progression of atherosclerotic plaque are scarce. Vascular smooth muscle cells (VSMCs) are important components of atherosclerotic plaques and oxidized low density lipoprotein (ox-LDL) is an important detrimental factor of atherosclerosis. The aim of this study is to elucidate the effect of TFPI-2 on smooth muscle cell proliferation and migration induced by ox-LDL. Methods: Retroviruses expressing human TFPI-2 were constructed. Cell proliferation was determined by CCK-8 assay. Cell apoptosis was analyzed by double staining of FITC-Annexin V and propidium iodide. Cell migration was studied through a Transwell chamber and with a scratch-wound assay. The matrix metalloproteinase-2 and −9 activities were analyzed by gelatin zymography. Phosphorylation of FAK was analyzed by western blot. Results: TFPI-2 over-expression of mRNA and protein was confirmed in infected cells. CCK-8 assay showed that TFPI-2 inhibit VSMCs proliferation induced by ox-LDL while without cytotoxicity to VSMCs. Transwell and scratch wound assay confirmed TFPI-2 over-expression can inhibit VSMC migration. Zymography assay showed that TFPI-2 can inhibit MMP-2, 9 activity induced by ox-LDL. Western blot assay showed TFPI-2 can inhibit cyclinD1 expression and FAK phosphorylation. Conclusion: TFPI-2 over-expression may strongly inhibit the proliferation and migration of VSMCs and suppresses MMP-2, 9 activity induced by ox-LDL, making it a promising candidate for treatment of atherosclerotic process.
tissue factor pathway inhibitor-2 (, TFPI-2), vascular smooth muscle cells (, VSMCs), proliferation migration atherosclerosis
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【期刊论文】Hypomethylated DSCR4 is a placenta-derived epigenetic marker for trisomy 21
马端, Yingying Du†, Jin Zhang†, Huijun Wang, Xiaoling Yan, Yingjun Yang, Lu Yang, Xin Luo, Yating Chen, Tao Duan* and Duan Ma, *
PRENATAL DIAGNOSIS Prenat Diagn 2011; 31: 207-214.,-0001,():
-1年11月30日
Background Trisomy 21 is the most common chromosomal aberration in live births. Some efforts have been made to develop noninvasive prenatal detection of trisomy 21 by using fetal DNA in maternal plasma. Due to the maternal DNA background, a distinguishable marker between maternal DNA and fetal DNA must be used, such as DNA methylation. The objective of this study was to search for fetal-specific methylation markers on chromosome 21. Methods We chose six genes highly or specifically expressed in placenta and screened the methylation status of these gene promoter regions by combination of methylated-DNA precipitation and methylation-sensitive restriction enzymes assay (COMPARE-MS). We further determined the methylation status of each CpG site within selected gene fragments by bisulfite sequencing. At last, we detected the placenta-derived methylation marker in the first-trimester maternal plasma by real-time methylation-specific PCR (MSP). Results Down syndrome (DS) critical region gene 4 (DSCR4) promoter region was found to be hypomethylated in placental tissues and densely methylated in maternal blood cells. Unmethylated DSCR4 (Down syndrome) sequence can be detected in the first-trimester maternal plasma. Conclusion DSCR4 promoter DNA is a candidate fetal epigenetic marker for noninvasive prenatal detection of trisomy 21. Copyright 2011 John Wiley & Sons, Ltd.
cell-free fetal DNA, DNA methylation, trisomy 21, noninvasive prenatal detection
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【期刊论文】The cell growth suppressor, mir-126, targets IRS-1
马端, Jin Zhang a, b, Ying-ying Du a, Yi-feng Lin a, Ya-ting Chen a, Lu Yang a, Hui-jun Wang b, Duan Ma a, *
Biochemical and Biophysical Research Communications 377 (2008) 136-140,-0001,():
-1年11月30日
miRNAs are a family of approximately 22-nuleotide-long noncoding RNAs involved in the formation and progress of tumors. Since traditional methods for the detection of miRNAs expression have many disadvantages, we developed a simple method called polyA RT PCR. With this method, we detected a series of miRNAs and found that mir-126 is one of the miRNAs underexpressed in breast cancer cells. Flow cytometry analysis showed that mir-126 inhibited cell cycle progression from G1/G0 to S. Further studies revealed that mir-126 targeted IRS-1 at the translation level. Knocking down of IRS-1 suppresses cell growth in HEK293 and breast cancer cell MCF-7, which recapitulates the effects of mir-126. In conclusion, we developed a simple method for high-throughput screening of miRNAs and found that mir-126, a cell growth suppressor, targets IRS-1.
microRNA, mir-126, Cell, Target genes, IRS-1, PolyA RT PCR
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