上皮膜蛋白1（lepithelial membrane protein 1,EMP1）基因位于第12号染色体长臂上[1],编码一种含4段跨膜区域的糖蛋白,参与涉及细胞周期调控、肿瘤形成、肿瘤细胞增殖及分化等多方面的细胞间信号传导通路[2].近年来国内外研究表明,EMP...上皮膜蛋白1（lepithelial membrane protein 1,EMP1）基因位于第12号染色体长臂上[1],编码一种含4段跨膜区域的糖蛋白,参与涉及细胞周期调控、肿瘤形成、肿瘤细胞增殖及分化等多方面的细胞间信号传导通路[2].近年来国内外研究表明,EMP1基因在胃癌、乳腺癌及子宫平滑肌瘤等许多肿瘤中均显著表达,并且在脑胶质瘤中也存在表达增高,同时与一些已证实的癌基因及相关分子关系密切.展开更多
目的:探讨上皮膜蛋白1基因(epithelial membrane protein 1,EMP1)在人脑胶质瘤中的表达及其与肿瘤恶性程度的相互关系。方法:肿瘤组织标本均取自上海长征医院神经外科2005至2006年部分手术病例,正常脑组织来源于捐献,分别采用逆转录酶...目的:探讨上皮膜蛋白1基因(epithelial membrane protein 1,EMP1)在人脑胶质瘤中的表达及其与肿瘤恶性程度的相互关系。方法:肿瘤组织标本均取自上海长征医院神经外科2005至2006年部分手术病例,正常脑组织来源于捐献,分别采用逆转录酶聚合酶链反应(RT-PCR)和免疫组化染色方法检测33例星形细胞瘤、3例少枝胶质细胞瘤、2例室管膜瘤以及7例正常脑组织标本中EMP1 mRNA及蛋白的表达。结果:EMP1无论在基因还是在蛋白表达水平上,在胶质瘤和正常脑组织中均有不同程度表达,但在星形细胞瘤中表达的含量明显高于正常脑组织,且在星形细胞瘤中含量随着其病理分级的增高而增高,在低级别(WHOⅠ～Ⅱ级)与高级别(Ⅲ～Ⅳ级)之间存在显著差异(蛋白P<0.05;基因P<0.01)。结论:基因EMP1在人脑胶质瘤中显著高表达,并且与星形细胞瘤的恶性程度密切相关。展开更多
AIM: To obtain human esophageal cancer cell EC9706 stably expressed epithelial membrane protein-1 (EMP-1) with integrated eukaryotic plasmid harboring the open reading frame (ORF) of human EMP-1, and then to study the...AIM: To obtain human esophageal cancer cell EC9706 stably expressed epithelial membrane protein-1 (EMP-1) with integrated eukaryotic plasmid harboring the open reading frame (ORF) of human EMP-1, and then to study the mechanism by which EMP-1 exerts its diverse cellular action on cell proliferation and altered gene profile by exploring the effect of EMP-1.METHODS: The authors first constructed pcDNA3.1/mychis expression vector harboring the ORF of EMP-1 and then transfected it into human esophageal carcinoma cell line EC9706. The positive clones were analyzed by Western blot and RT-PCR. Moreover, the cell growth curve was observed and the cell cycle was checked by FACS technique. Using cDNA microarray technology, the authors compared the gene expression pattern in positive clones with control. To confirm the gene expression profile, semi-quantitative RT-PCR was carried out for 4 of the randomly picked differentially expressed genes. For those differentially expressed genes,classification was performed according to their function and cellular component.RESULTS: Human EMP-1 gene can be stably expressed in ECg706 cell line transfected with human EMP-1. The authors found the cell growth decreased, among which S phase was arrested and G1 phase was prolonged in the transfected positive clones. By cDNA microarray analysis, 35 genes showed an over 2.0 fold change in expression level after transfection, with 28 genes being consistently up-regulated and 7 genes being down-regulated. Among the classified genes, almost half of the induced genes (13 out of 28 genes) were related to cell signaling, cell communication and particularly to adhesion.CONCLUSION: Overexpression of human EMP-1 gene can inhibit the proliferation of EC9706 cell with S phase arrested and G1 phase prolonged. The cDNA microarray analysis suggested that EMP-1 may be one of regulators involved incell signaling, cell communication and adhesion regulators.展开更多
文摘目的:探讨上皮膜蛋白1(epithelial membrane protejn-1,EMP1)在人结直肠癌(colorectal carcinoma,CRC)组织中的表达水平及其过表达对CRC SW-480细胞增殖、凋亡和侵袭的影响。方法:免疫组织化学、Western blotting方法检测63例CRC组织及31例癌旁组织中EMP1的表达水平,分析EMP1表达与CRC临床病理参数的关系。慢病毒介导将pLenti6-EMP1质粒转染SW-480细胞,建立EMP1过表达细胞,转染plenti6/V5-DEST空白质粒为对照。Real-time PCR及Western blotting检测转染后SW-480细胞株中EMP1的表达,MTT、流式细胞术及Transwell实验分别检测EMP1过表达对SW-480细胞增殖、凋亡及侵袭能力的影响。结果:EMP1蛋白在人CRC组织的表达显著低于癌旁组织(0.257±0.022 vs 0.863±0.086,P<0.05),并且其表达水平在不同T分期、有无淋巴结转移、临床分期以及组织分级组间表达有显著差异(P<0.05),而与患者年龄、性别、肿瘤部位无关(P>0.05)。成功构建EMP1过表达的LeEMP1细胞。与LeEmpty细胞相比,LeEMP1细胞的增殖能力显著下降[(60.94±4.04)%vs(100.00±0.00)%,P<0.05],凋亡率显著升高[(12.10±1.30)%vs(3.10±0.60)%,P<0.05]、侵袭转移能力显著降低[穿膜细胞数:(87.00±12.00)vs(178.00±21.00)个,P<0.05]。LeEMP1细胞Caspase-9表达显著高于LeEmpty细胞(0.764±0.073 vs 0.231±0.029,P<0.05),VEGFC表达显著降低(0.185±0.022 vs 0.663±0.065,P<0.05)。结论:人CRC组织中EMP1蛋白表达明显减低,过表达EMP1能够抑制CRC SW-480细胞的恶性生物学行为,其可能通过调控Caspase-9和VEGFC的表达来发挥作用。
文摘上皮膜蛋白1（lepithelial membrane protein 1,EMP1）基因位于第12号染色体长臂上[1],编码一种含4段跨膜区域的糖蛋白,参与涉及细胞周期调控、肿瘤形成、肿瘤细胞增殖及分化等多方面的细胞间信号传导通路[2].近年来国内外研究表明,EMP1基因在胃癌、乳腺癌及子宫平滑肌瘤等许多肿瘤中均显著表达,并且在脑胶质瘤中也存在表达增高,同时与一些已证实的癌基因及相关分子关系密切.
基金国家高科技研究发展(863)计划项目(No.2005AA001070)Supported by the National High Technology Research and Development Program of China(No.2005AA001070)
文摘目的:探讨上皮膜蛋白1基因(epithelial membrane protein 1,EMP1)在人脑胶质瘤中的表达及其与肿瘤恶性程度的相互关系。方法:肿瘤组织标本均取自上海长征医院神经外科2005至2006年部分手术病例,正常脑组织来源于捐献,分别采用逆转录酶聚合酶链反应(RT-PCR)和免疫组化染色方法检测33例星形细胞瘤、3例少枝胶质细胞瘤、2例室管膜瘤以及7例正常脑组织标本中EMP1 mRNA及蛋白的表达。结果:EMP1无论在基因还是在蛋白表达水平上,在胶质瘤和正常脑组织中均有不同程度表达,但在星形细胞瘤中表达的含量明显高于正常脑组织,且在星形细胞瘤中含量随着其病理分级的增高而增高,在低级别(WHOⅠ～Ⅱ级)与高级别(Ⅲ～Ⅳ级)之间存在显著差异(蛋白P<0.05;基因P<0.01)。结论:基因EMP1在人脑胶质瘤中显著高表达,并且与星形细胞瘤的恶性程度密切相关。
基金China Key Program on Basic Research(G1998051021)the Chinese Hi-Tech R&D program(2001AA231310)
文摘AIM: To obtain human esophageal cancer cell EC9706 stably expressed epithelial membrane protein-1 (EMP-1) with integrated eukaryotic plasmid harboring the open reading frame (ORF) of human EMP-1, and then to study the mechanism by which EMP-1 exerts its diverse cellular action on cell proliferation and altered gene profile by exploring the effect of EMP-1.METHODS: The authors first constructed pcDNA3.1/mychis expression vector harboring the ORF of EMP-1 and then transfected it into human esophageal carcinoma cell line EC9706. The positive clones were analyzed by Western blot and RT-PCR. Moreover, the cell growth curve was observed and the cell cycle was checked by FACS technique. Using cDNA microarray technology, the authors compared the gene expression pattern in positive clones with control. To confirm the gene expression profile, semi-quantitative RT-PCR was carried out for 4 of the randomly picked differentially expressed genes. For those differentially expressed genes,classification was performed according to their function and cellular component.RESULTS: Human EMP-1 gene can be stably expressed in ECg706 cell line transfected with human EMP-1. The authors found the cell growth decreased, among which S phase was arrested and G1 phase was prolonged in the transfected positive clones. By cDNA microarray analysis, 35 genes showed an over 2.0 fold change in expression level after transfection, with 28 genes being consistently up-regulated and 7 genes being down-regulated. Among the classified genes, almost half of the induced genes (13 out of 28 genes) were related to cell signaling, cell communication and particularly to adhesion.CONCLUSION: Overexpression of human EMP-1 gene can inhibit the proliferation of EC9706 cell with S phase arrested and G1 phase prolonged. The cDNA microarray analysis suggested that EMP-1 may be one of regulators involved incell signaling, cell communication and adhesion regulators.
