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Cripto-1基因沉默对人肝癌MHCC-97H细胞侵袭、迁移能力的影响
  • ISSN号:1000-4718
  • 期刊名称:《中国病理生理杂志》
  • 分类:R730.23[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:中山大学附属第一医院肝外科,广东广州510080
  • 相关基金:国家自然科学基金资助项目(No.81472254)
中文摘要:

目的:探讨Cripto-1蛋白在人肝癌组织和肝癌细胞系中的表达,研究沉默Cripto-1基因后对肝癌细胞MHCC-97H侵袭和迁移能力的影响及可能的机制。方法:采用Western blot检测11对肝癌组织及相应的癌旁组织和不同肝癌细胞株中Cripto-1蛋白的表达;收集80例肝癌石蜡标本,免疫组织化学方法检测Cripto-1蛋白的表达情况;Cripto-1 siRNA转染肝癌细胞MHCC-97H细胞后,real-time PCR和Western blot分别检测转染效率;Transwell迁移和侵袭实验分别检测细胞的迁移和侵袭能力;Western blot分别检测基质金属蛋白酶(MMP)-2、MMP-9蛋白的表达。结果:Western blot检测结果显示肝癌组织中Cripto-1蛋白的表达明显高于对应的癌旁组织,Cripto-1蛋白在各肝癌细胞系中均有表达,且表达量高于对照细胞,其中高侵袭性肝癌细胞MHCC-97H表达量最高;免疫组织化学结果显示Cripto-1蛋白在88.7%的肝癌组织中表达;Cripto-1 siRNA转染MHCC-97H细胞后能显著降低Cripto-1mRNA和蛋白的表达水平,并且能降低其侵袭和迁移能力;Western blot结果表明,Cripto-1基因沉默后能抑制MMP-2和MMP-9蛋白的表达。结论:Cripto-1在肝细胞癌中的高表达可能与肝癌的发生发展密切相关,下调Cripto-1的表达可以抑制肝癌细胞侵袭和迁移能力,其机制可能与下调MMP-2、MMP-9的表达有关。

英文摘要:

AIM: To investigate the expression of Cripto-1 in hepatocellular carcinoma( HCC) and the effects of Cripto-1 gene silencing on the invasion and migration of MHCC-97 H cells. METHODS: The protein expression of Cripto-1 in the tumor tissues,paracancerous tissues and different HCC cell lines was detected by Western blot. The hepatocellular carcinoma paraffin-embedded specimens( n = 80) were collected. The protein expression of Cripto-1 was detected by immunohistochemical method. The Cripto-1 siRNA or control siRNA was transfected into the MHCC-97 H cells. The expression of Cripto-1 at mRNA and protein levels was detected by real-time PCR and Western blot,respectively. Transwell migration and invasion experiments were performed to measure the migration and invasion abilities. Finally,the protein expression of matrix metalloprotein-ase( MMP)-2 and MMP-9 in the MHCC-97 H cells transfected with Cripto-1 siRNA was detected by Western blot. RESULTS: The protein expression of Cripto-1 in the tumor tissues was significantly higher than that in the paracancerous tissues. Meanwhile,the protein expression of Cripto-1 in the HCC cell lines( highest in MHCC-97 H cells) was higher than that in the control cell line. The results of immunohistochemistry show the positive rate of Cripto-1 in HCC was 88. 7%. Compared with control group,the expression of Cripto-1 at mRNA and protein levels was notably down-regulated in the MHCC-97 H cells transfected with Cripto-1 siRNA,so was the expression of MMP-2 and MMP-9. In addition,the migration and invasion abilities of MHCC-97 H cells transfected with Cripto-1 siRNA were decreased. CONCLUSION: Over-expression of Cripto-1 may be closely associated with the occurrence and development of HCC. Cripto-1gene silencing decreases the migration and invasion abilities of MHCC-97 H cells,which may act via inhibiting the expression of MMP-2 and MMP9.

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期刊信息
  • 《中国病理生理杂志》
  • 中国科技核心期刊
  • 主管单位:中国科学技术协会
  • 主办单位:中国病理生理学会
  • 主编:陆大祥
  • 地址:广东省广州市黄埔大道西601号
  • 邮编:510632
  • 邮箱:obsbjbb@jnu.edu.cn
  • 电话:020-85220269
  • 国际标准刊号:ISSN:1000-4718
  • 国内统一刊号:ISSN:44-1187/R
  • 邮发代号:46-98
  • 获奖情况:
  • 1997-2000年连续获得中国科协优秀基础性和高科技...,1992、1996、2000、2004、2008年,连续五次入选中...,2008-2010年,连续三年荣获“百种中国杰出学术期...,2010年获广东省期刊最高奖——品牌期刊奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:37010