期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

肝癌抑制基因-1与转运功能相关的451bp序列区的确定 被引量：1

Establishment of 451 bp fragment that functionally related to transportation in hepatocellular carcinoma supressor gene-1(HCCS1)
下载PDF
导出
摘要 背景与目的:肝癌抑制基因-1(HCCS1)是一种潜在的肝癌抑制基因,并且在细胞内蛋白分拣运输中也发挥着重要作用,其抑癌作用有可能是通过其蛋白转运的功能而发挥的,本文以寻找HCCS1序列中与转运相关的最小功能序列区域为目的。方法:通过亚克隆技术,构建以pEGFP-C2为载体的含不同长度HCCS1cDNA片段的亚克隆,将构建的亚克隆转染子宫颈癌HeLa细胞,通过免疫荧光共聚焦显微镜观察不同长度HCCS1蛋白的分布,以及与6-磷酸甘露糖受体(M6PR)的共定位。结果:成功构建了以pEGFP-C2为载体的10个含有不同长度HCCS1片段的亚克隆;HCCS1cDNA从3'端向5'端逐渐缺失的片段中:2100bp片段至778bp片段编码的不同长度HCCS1蛋白呈颗粒状分布于核周的胞质内,其中2100bp片段至1571bp片段编码的不同长度HCCS1蛋白均呈颗粒状、极性分布于核周的胞质内,且与M6PR有共定位;而1120bp片段至778bp片段编码的不同长度HCCS1蛋白虽然也呈颗粒状分布于核周,但极性分布消失,且与M6PR共定位消失;HCCS1cDNA片段678bp片段至339bp片段编码的不同长度HCCS1蛋白呈散点状弥散分布于胞质以及胞核内。结论:确定了HCCS1cDNA1571bp片段的3'端451bp的序列为与HCCS1转运功能相关的最小区域范围。 Background and purpose： Hepatocellular carcinoma suppressor gene-1 （HCCS1） is a potential hepatocellular carcinoma supressor gene, and it also plays an important role in sorting of some cytoplasmic proteins. Its carcinoma suppressor function may be related to its sorting function. So it is important to identify the minimum region of functional sequence in HCCS1 that related to the transportation. Methods： The expression vectors containing various lengths of HCCS1 gene were constructed and transfected into HeLa cells mediated by liposomes. The localizations of different HCCS1 frag- ments and the colocalizations of M6PR with various truncated HCCS1 were determined by laser scanning confocal microscopy, respectively. Results： 10 different expression vectors containing various lengths of HCCS1 gene were successfully constructed, it had been shown that the polar localization of HCCS1 protein and the co-localizations with M6PR disappeared when HCCS1 gene was truncated to 1 120 bp from the 3＇ end by laser scanning confocal microscopy. Conclusions： We identified the minimum localization of the HCCS1 functional sequence that related to the transportation.
作者 孔伟 甘愉 胡晶莹 朱明洁 赵仰星 赵新泰
机构地区 复旦大学上海医学院 上海市肿瘤研究所细胞与分子免疫学实验室
出处 《中国癌症杂志》 CAS CSCD 2008年第1期1-5,共5页 China Oncology
基金 国家自然科学基金重点资助项目(No.30330350) 上海市科委资助项目(No.04XD14015)
关键词 肝癌抑制基因 功能序列 转运 免疫荧光共聚焦 hepatocellular carcinoma supressor gene functional sequence transportation confocal immuno-fluorescence microscopy
分类号 R735.7 [医药卫生—肿瘤]
  • 相关文献

参考文献10

  • 1Zhao XT, Li JJ, He YH, et al. A novel growth suppressor gene on chromosome 17p13.3 with a high frequency of mutation in human hepatocellular carcinoma [ J ]. Cancer Res, 2001,61 ( 20 ) : 7383-7387.
  • 2Liewen H, Ivo Meinhold-Heerlein, Vasco Oliveira, et al. Characterization of the human GARP ( Golgi associated retrograde protein) complex[J]. Exp Cell Res,2005,306( 1 ) :24-34.
  • 3Conibear E,Stevens TH. Vps52p, Vps53p, and Vps54p form a novel multisubunit complex required for protein sorting at the yeast late Golgi [ J ]. Mol Biol Cell ,2000,11 ( 1 ) :305-323.
  • 4Conibear E, Stevens TH. Studying yeast vacuoles [ J ]. Methods Enzymol,2002 ,351:408-432.
  • 5Andag U, Neumann T, Schmitt HD. The coatomer-interacting protein Dsll p is required for Golgi-to-endoplasmic reticulum retrieval in yeast[ J]. Biol Chem,2001,276(42) :39150-39160.
  • 6Conibear E, Cleck JN, Stevens TH. Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1 p [ J ]. Mol. Biol. Cell,2003,14 (4) : 1610-1623.
  • 7Nicole R, Qwennwvillw NR, Chao TY, et al. Domains within the GARP subunit Vps54 confer separate functions in complex assembly and early endosome recognition [ J ]. Mol Biol Cell,2006, 17(4) :1859-1870.
  • 8Wiley SH, Burke PM. Regulation of receptor tyrosine kinase signaling by endocytic trafficking[J]. Traffic,2001,2( 1 ) :12-18.
  • 9Piao Z, Park C, Park JH, et al. Allelotype analysis of hepatocellular carcinoma. [J]. Cancer,1998,75(8) :29-33.
  • 10Charroux B, Shevchenko A, Mann M, et al. Gemin4, A novel component of the SMN complex that is found in both gems and nucleoli[J]. Cell Biol,2000,148(6) :1177-1186.

同被引文献11

  • 1Liu X Y,Gan Y,Zhao X,et al.Adenovirus-mediated HCCS1 overexpression elicits a potent antitumor efficacy on human colorectal cancer and hepatoma cells both in vitro and in vivo[J].Cancer Gene Therapy,2008,15:808-816.
  • 2赵新泰,李锦军,何英华,等.肝癌中具有高突变率位于染色体17p13.3区的新抑癌基因.癌症研究,2001,.
  • 3Liewen H,lvo Meinhold-Heerlein,Vasco Oliveira,et al.Characterization of the human GARP(GO ls～associated retrograde protein) complex[J].Exp Cell Res,2005,306(1):24-34.
  • 4Piao Z,Park C,Park J H,et al.Allelotype analysis of hepatocellular carcinoma[J].Cancer,1998,75(8):29-33.
  • 5Charroux B,Shevehenko A,Mann M,et al.Gemin4,A novel component of the SMN complex that is found in both gems and nueleoli[J].Cell Biol,2000,148(6):1177-1186.
  • 6Conibear E,Stevens T H.Vps52p,Vps53p,and Vps54p form a novel muhisubunit complex required for protein sorting at the yeastlateGOM[J].Mol Biol Cell,2000,11(1):305-323.
  • 7Conibear E,Stevens T H.Studying yeast vacuoles[J].Methods Enzymol,2002,351:408-432.
  • 8Andag U,Neumann T,Sehmitt H D.The coatomer-interacting protein Dsll P is required for GO lsl-to-endoplasmie retieulum re-trieval in yeast[J].Biol Chem,2001,276(42):39150-39160.
  • 9Conibear E,Cleek J N,Stevens T H.Vps51P mediates the association of the GARP(Vps52/53/54)complex with the late Golsl t-SNARE TlslP[J].Mol Biol Cell,2003,14(4):1610-1623.
  • 10Nieole R,Qwennwvillw N R,Chao T Y,et al.Domains within the GARP subunit Vps54 confer separate functions in complex as-sembly and early endosome recognition[J].Mol Biol Cell,2006,17(4):1859-1870.

引证文献1

中国癌症杂志
2008年 第1期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部