[Construction of adenovirus vector expressing TIP30 and its tumor suppressive effect in vitro and in vivo].
OBJECTIVE
To construct an adenovirus vector expressing TIP30 gene (Ad-TIP30) and investigate its tumor suppressive effect in vitro and in vivo.


METHODS
Ad-Easy system was used to construct Ad-TIP30 by recombination in E. coli. The virus was packaged in 293 cells and subsequently identified valid. Human HCC (hepatocellular carcinoma) cell lines HepG(2) (p53-wt), PLC/PRL/5 (p53-mut), and osteosarcoma cell line Saos-2 (p53-null) with different p53 genotype were infected with Ad-TIP30 and control virus with Ad-GFP, respectively. The tumor suppressive effect of TIP30 in vitro was examined by trypan blue exclusion method. The expression level of p53 was determined by RT-PCR before and after Ad-TIP30 infection. The in vivo tumor suppressive effect was detected in nude mice with human HCC xenograft.


RESULTS
The expression of TIP30 significantly inhibited the in vitro proliferation of tumor cells, among which HepG(2) with wild type p53 gene was most susceptible to Ad-TIP30 induced growth inhibition. The expression of p53 was significantly up-regulated in HepG(2) after Ad-TIP30 infection as determined by RT-PCR. The growth in nude mice of HCC infected with Ad-TIP30 was significantly inhibited with an inhibition rate of 62.9%.


CONCLUSIONS
The expression of TIP30 could inhibit the proliferation of tumor cell lines through both p53-dependent and p53-independent pathways, and may be used as a potential tool for cancer therapy.
Zhang
Xia
X
The Second Military Medical University Cancer Institute, Eastern Hepatobiliary Surgical Hospital, Shanghai 200433, China.
Zhao
Jian
J
Li
Xiao-dong
XD
Yuan
Chang-ting
CT
Wang
Hua-qing
HQ
Wu
Meng-chao
MC
Xiao
Hua
H
Guo
Ya-jun
YJ
D000123:Acetyltransferases / Q000235:genetics*; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002455:Cell Division; D016158:Genes, p53; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D051379:Mice; D009374:Neoplasms, Experimental / Q000235:genetics / Q000628:therapy*; D014157:Transcription Factors / Q000235:genetics*
D014157:Transcription Factors; D000123:Acetyltransferases; C111246:HTATIP2 protein, human