Construction and Expression of Hepatocellular Carcinoma-Specific Expressing Eukaryotic Vector for Anti-Angiogenesis Therapy
- VernacularTitle:肝癌特异性血管抑制疗法的真核表达载体的构建及其表达
- Author:
Hongyong XU
;
Li XU
;
Jianhong GAO
;
Kaizong LI
;
Kefeng DOU
- Publication Type:Journal Article
- Keywords:
Hepatocellular carcinoma Angiostatin ?-Fetoprotein enhancer
- From:
Chinese Journal of Bases and Clinics in General Surgery
2004;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a mammalian vector encoding angiostatin kringle 5 (K5) under the control of ?-fetoprotein (AFP) enhancer and albumin promoter, and to observe the expression of angiostatin by introducting angiostatin gene into hepatocellular carcinoma cells through gene transfection. Methods Angiostatin cDNA was amplified from normal human eukaryotic cells by using RT-PCR. Meanwhile, AFP enhancer and albumin promoter sequences were directed cloned and were inserted into vector pcDNA3.1. The recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was constructed, which contained the angiostatin K5 cDNA sequence that was under the control of the AFP enhancer and promoter. Angiostatin K5 cDNA was introduced into human AFP positive hepatocellular carcinoma cell lines with the transfected cultured cells that were mediated with Lipofectamine 2000. The expression of angiostatin K5 was analyzed by Western blot and the protein was dectected with anti-His antibody. Results The 500-base pair of angiostatin K5 was in accordance with the expected sequence and the recombinant vector of pcDNA3.1-AFAB-angiostatin K5-His was also confirmed as the anticipated sequence. The expression of angiostatin K5 in AFP positive hepatocellular carcinoma cells was detected both by SDS-PAGE and Western blot. Conclusion Efficient construction and expression of angiostatin K5 to AFP positive cells make it possible for antiangiogenesis therapy of human hepatocellular carcinomas, which may provide a promising approach.
