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p62增加HBsAg+ HepG2细胞DRibbles中HBsAg含量的研究
  • ISSN号:1671-6264
  • 期刊名称:《东南大学学报:医学版》
  • 时间:0
  • 分类:R392.11[医药卫生—免疫学;医药卫生—基础医学] R-33
  • 作者机构:[1]东南大学医学院病原生物与免疫学系,江苏南京210009
  • 相关基金:国家自然科学基金资助项目(30771999 30972783); 江苏省自然科学基金资助项目(BK20082096)
中文摘要:

目的:探讨过表达p62能否提高自噬小体中泛素化蛋白的水平。方法:构建pIRES2-EGFP-HBsAg重组质粒和pIRES2-EGFP-Ub-HBsAg重组质粒;脂质体法分HBsAg、Ub-HBsAg、Ub-HBsAg+p62组3组转染HepG2细胞,在相同细胞数量和转染效率下,经万珂、雷帕霉素和氯化铵处理转染细胞并提取DRibbles,ELISA法检测其中HBsAg含量。结果:万珂、雷帕霉素和氯化铵处理后,Ub-HBsAg+p62组HepG2细胞DRibbles中HBsAg显著增高,而且明显高于药物处理的其他两组。结论:当蛋白酶体被抑制时,过表达p62蛋白促进泛素化蛋白(HBsAg)进入自噬小体,阻断自噬降解途径提取的DRibbles能够募集到更多的泛素化蛋白(HBsAg)。

英文摘要:

Objective:To approach whether over-expressed p62 could aggregate more ubiquitinated proteins in DRibbles.Methods:Recombinant pIRES2-EGFP vector containing the HBsAg or UbHBsAg gene was constructed and transfected into HepG2 cell line respectively and pIRES2-EGFP-UbHBsAg and pIRES2-EGFP-p62 were cotransfected into HepG2 cell line.Then the transfected cells were cultured in the medium including velcade,rapamycin and ammonium chloride.After 16 hours,we extracted and lysed DRibbles from the medium and detected HBsAg by ELISA.Results:HBsAg in DRibbles extracted from HepG2 that was co-transfected with pIRES2-EGFPUbHBsAg and pIRES2-EGFP-p62 vectors rapidly increased when the cells were cultured with the drugs,and was much higher than the other groups which had been under the same treating.Conclusion:Over-expression of p62 aggregates more ubiquitinated proteins(HBsAg) in the DRibbles extracted from HepG2 cells as the proteasome pathway is blocked by the drugs.

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期刊信息
  • 《东南大学学报:医学版》
  • 中国科技核心期刊
  • 主管单位:教育部
  • 主办单位:东南大学
  • 主编:唐萌
  • 地址:南京市鼓楼区丁家桥87号
  • 邮编:210009
  • 邮箱:bjb@pub.seu.edu.cn
  • 电话:025-83272481
  • 国际标准刊号:ISSN:1671-6264
  • 国内统一刊号:ISSN:32-1647/R
  • 邮发代号:28-265
  • 获奖情况:
  • 全国优秀学报,铁道部优秀期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引,中国中国科技核心期刊,中国北大核心期刊(2014版)
  • 被引量:8241