Reconstruction of adenovirus vector contained canstatin gene and hTERT gene core promoter and its anti-tumor effect in human ovarian cancer
10.3760/cma.j.issn.0529-567x.2009.03.014
- VernacularTitle:含hTERT基因核心启动子和canstatin基因重组腺病毒载体的构建及其对卵巢上皮性癌的抑制作用
- Author:
Baoju ZHU
;
Yuhuan QIAO
;
Yin LI
- Publication Type:Journal Article
- Keywords:
Ovarian neoplasms;
Telomerase;
Promoter regions ( genetics );
Peptidefragments;
Collagen type Ⅳ;
Angiogenesis inhibitors;
Adenoviridae
- From:
Chinese Journal of Obstetrics and Gynecology
2009;44(3):214-218
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study anti-tumor effects on human ovarian cancer xenografted tumors in mice by constructing adenoviral expression vector containing canstatin gene and hTERT gene core promoter (AdhTERT-Can). Methods AdhTERT-Can vector was constructed and identified by means of enzyme cutting, electrophoresis and sequencing. Then, transfected into HO8910PM cells by means of lipofectamine and confirmed by green fluorescence protein(GFP) expression under laser confocus microscope. The mRNA expression of canstatin gene was tested by RT-PCR. Human ovarian cancer xenografted tumor models in nude mice were established and randomly divided into AdhTERT-Can group received viral supematant solution of AdhTERT-Can by tail vein injection, Ad-Can groups received viral supernatant solution of Ad-Can by tail vein injection, and control groups received phosphate buffer solution (PBS). The volume of tumors were measured and compared in each group to evaluated the anti-tumor efficacy. Results All the constructed vectors of AdhTERT-Can were verified by enzymed digestion. There were green fluorescence from 60% of HO8910PM cells transfected by AdhTERT-Can under laser confocus microscope, and the mRNA expression of canstafin gene in HO8910PM cells were also verified by RT-PCR The growth of tumor in AdhTERT-Can group was significantly inhibited compared with those in Ad-Can groups and control groups from the 8th day (P <0.01 ). However, there were not significant difference between Ad-Can group and PBS group (P> 0.05). On the 30th day, the tumors showed liquefaction necrosis and cystic degeneration in each group, especially in AdhTERT-Can group. Conclusions The recombinant adenovirus vector of AdhTERT-Can has been constructed successfully and could steady express in ovarian cancer cell lines HO8910PM. The results shown that it could inhibit significantly the growth of human ovarian cancer xenografted tumors in mice and shown to be the target of gene therapy.
