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黏附素5胞外区克隆、转染及抗人乳腺癌细胞生长的研究
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作者 石小玉 李文林 +3 位作者 熊丽霞 张际青 熊建军 李红 《中国病理生理杂志》 CAS CSCD 北大核心 2007年第9期1710-1715,共6页
目的:克隆和转染黏附素5胞外区,并研究其对人乳腺癌MDA-MB435细胞株生长的影响。方法:RT-PCR技术克隆黏附素5胞外区(称为CED1-4),将其插入pMSCV质粒载体,在大肠杆菌XL-blue扩增,提取和纯化pMSCV-CED1-4,酶切、电泳和测序检测CED1-4序列... 目的:克隆和转染黏附素5胞外区,并研究其对人乳腺癌MDA-MB435细胞株生长的影响。方法:RT-PCR技术克隆黏附素5胞外区(称为CED1-4),将其插入pMSCV质粒载体,在大肠杆菌XL-blue扩增,提取和纯化pMSCV-CED1-4,酶切、电泳和测序检测CED1-4序列。CED1-4基因转染MDA-MB435细胞株,RT-PCR和Western blotting检测MDA-MB435细胞表达CED1-4。细胞增殖实验和乳腺癌裸小鼠致瘤实验检测CED1-4对MDA-MB435细胞体内外生长的影响。结果:构建了重组体pMSCV-CED1-4,电泳显示CED1-4条带在1636bp-1018bp区间,测序显示CED1-4基因长1452bp,编码484氨基酸。经PCR和Western blotting证实,CED1-4基因转染的MDA-MB435细胞在mRNA和蛋白质水平表达CED1-4。细胞增殖实验结果表明,实验组MDA-MB435细胞增殖低于实验对照组和空白对照组(P<0.05)。乳腺癌裸小鼠致瘤实验显示,实验组移植瘤平均体积和重量低于实验对照组和空白对照组(P<0.05)。结论:黏附素5胞外区CED1-4能在体内外抑制人乳腺癌MDA-MB435细胞株的生长。 展开更多
关键词 钙粘着糖蛋白类 克隆 乳腺肿瘤 基因转染
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人可溶性粘附素5截短体的克隆、表达及生物活性测定
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作者 李文林 石小玉 +3 位作者 熊丽霞 王志刚 周莹 李红 《中国免疫学杂志》 CAS CSCD 北大核心 2008年第1期62-64,70,共4页
目的:克隆和转染人可溶性粘附素5截短体(sCadherin5△),并检测其生物活性。方法:RT-PCR扩增Cadher-in5△cDNA,并克隆入pMSCV逆转录病毒载体,构建pMSCV/sCadherin5△,转化感受态大肠杆菌XL-blue菌株,提取、纯化和酶切质粒,测序分析sCadhe... 目的:克隆和转染人可溶性粘附素5截短体(sCadherin5△),并检测其生物活性。方法:RT-PCR扩增Cadher-in5△cDNA,并克隆入pMSCV逆转录病毒载体,构建pMSCV/sCadherin5△,转化感受态大肠杆菌XL-blue菌株,提取、纯化和酶切质粒,测序分析sCadherin5△,转染NIH3T3细胞,mRNA和蛋白质水平检测NIH3T3细胞表达和分泌sCadherin5△,MTT方法检测sCadherin5△生物活性。结果:PCR产物约为1128bp,构建了pMSCV/sCadherin5△质粒载体,序列测定的结果与GeneBank中Cadherin5胞膜外区121bp至1248bp之间的序列一致。pMSCV/sCadherin5△转染的NIH3T3细胞表达和分泌sCadherin5△,sCadherin5△抑制HUVEC细胞体外增殖。结论:克隆、表达和分泌了具有生物活性的人sCadherin5△。 展开更多
关键词 粘附素5 克隆 逆转录病毒载体 NIH3T3细胞 HUVEC细胞
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CONTROL OF ANGIOGENESIS BY INHIBITOR OF PHOSPHOLIPASE A_2
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作者 陈文明 李利红 +4 位作者 朱嘉芷 刘晋玮 Soria Jeannette Soria Claudine Yedgar Saul 《Chinese Medical Sciences Journal》 CAS CSCD 2004年第1期6-12,共7页
Objective To investigate the potential effects of angiogenic process by secretory phospholipase A2 (sPLA2)inhibitor-HyPE(linking N-derivatized phosphatidyl-ethanolamine to hyaluronic acid)on human bone marrow endothel... Objective To investigate the potential effects of angiogenic process by secretory phospholipase A2 (sPLA2)inhibitor-HyPE(linking N-derivatized phosphatidyl-ethanolamine to hyaluronic acid)on human bone marrow endothelial cell line(HBME-1). Methods In order to examine the suppressing effects of HyPE on HBME-1 proliferation, migration, and capillary-like tube formation, HBME-1 were activated by angiogenic factor, specifically by basic fibroblast growth factor(b-FGF), vascular endothelial growth factor(VEGF),and oncostatin M(OSM)(at a final concentration of 25, 20, and 2.5 ng/mL, respectively), then HBME-1 proliferation, migration, and tube forma-tion were studied in the absence or presence of HyPE. HBME-1 tube formation was specially analyzed in fibrin gel. Results HyPE effectively inhibited HBME-1 proliferation and migration as a dose-dependent manner, whatever HBME-1 were grown in the control culture medium or stimulated with b-FGF, VEGF, or OSM. In fibrin, the formations of HBME-1 derived tube-like structures were enhanced by all angiogenic factors, but these were strongly suppressed by HyPE. Conclusions The results support the involvement of sPLA2 in angiogenesis. It is proposed that sPLA2 inhibitor introduces a novel approach in the control of cancer development. 展开更多
关键词 ANGIOGENESIS phospholipase A_2 inhibitor endothelial cell line
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