Related Experiment Video
Updated: Aug 25, 2026
Live Cell Imaging of the TGF- β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System
Published on: July 30, 2018
[Construction of adenovirus vector expressing TIP30 and its tumor suppressive effect in vitro and in vivo]
Xia Zhang1, Jian Zhao, Xiao-dong Li
1The Second Military Medical University Cancer Institute, Eastern Hepatobiliary Surgical Hospital, Shanghai 200433, China.
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%.
Conclusion:
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.
