Colorectal Cancer
Copyright ©The Author(s) 2003. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 15, 2003; 9(5): 965-969
Published online May 15, 2003. doi: 10.3748/wjg.v9.i5.965
Tumor necrosis factor-related apoptosis-inducing ligand gene on human colorectal cancer cell line HT29
Xiang-Ming Xu, Chao He, Xiao-Tong Hu, Bing-Liang Fang
Xiang-Ming Xu, Department of Colorectal Surgery of the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China
Chao He, Xiao-Tong Hu, Clinical Research Institute of Sir Run Run Shaw Hospital, Zhejiang University, Hangzhou 310016, Zhejiang Province, China
Bing-Liang Fang, Department of Thoracic and Cardiovascular Surgery, the University of Texas MD. Anderson Cancer Center, Box109, 1515 Holcombe Boulevard, Houston, TX77030, Texas, USA
Author contributions: All authors contributed equally to the work.
Supported by the Scientific Committee Foundation of Zhejiang Province, No. 001103163
Correspondence to: Chao He, Clinical Research Institute of Sir Run Run Shaw Hospital, Zhejiang University, Hangzhou 310016, Zhejiang Province, China. drhe@zju.edu.cn
Telephone: +86-571-86048962
Received: October 9, 2002
Revised: October 23, 2002
Accepted: November 6, 2002
Published online: May 15, 2003
Abstract

AIM: To evaluate the therapeutic efficiency of Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) gene on human colorectal cancer cell line HT29.

METHODS: Human embryonal kidney cells transformed by introducing sheared fragments of Ad5 DNA (293 cell) were used for amplification of adenoviral vectors: Ad/GT-TRAIL, Ad/GT-Bax, Ad/GT-LacZ and Ad/PGK-GV16. Human colorectal cancer cell line HT29 was transfected with binary adenovirus-mediated TRAIL gene. Bax gene was used as positive control, LacZ gene was used as the vector control, and cells treated with PBS only were used as a mock control. The morphological changes, cell growth and apoptosis were measured by reversmicroscope, MTT method and flow cytometry.

RESULTS: All adenoviral vectors titer determined by optical absorbency at A260nm were 1 × 1010 viral particle/ml(vp/ml). Obviously morphological changes of HT29 cells were observed when infected with Ad/GT-TRAIL, and these changes were much more obviously when Ad/PGK-GV16 was coinfected. The cell suppression percentage and the percentage of apoptotic cells were 52.5% and 16.5% respectively when infected with Ad/GT-TRAIL alone, while combining with Ad/PGK-GV16, the growth of HT29 was suppressed by 85.2% and the percentage of apoptotic cells was 35.9%. It showed a significantly enhanced therapeutic efficiency with binary system (P < 0.05).

CONCLUSION: A binary adenoviral vector system provides an effective approach to amplify viral vectors that express potentially toxic gene, TRAIL. Ad/GT-TRAIL showed a significantly enhanced therapeutic efficiency for HT29 when coinfected with Ad/PGK-GV16. Ad/GT-TRAIL could induce apoptosis of HT29 and inhibit its growth.

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