[Frontiers in Bioscience E4, 2389-2395, June 1, 2012]

Adenovirus KH901 promotes 5-FU antitumor efficacy and S phase in LoVo cells

Wei Peng1, Jin Li1, XG Yin2, JF Xu1, LZ Cheng1

1Department of Medical Oncology, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University,shanghai 200032, China, 2Chengdu Kanghong Biotechnology Co., Ltd, 36 Xishu Rd, Chengdu, Sichuan 610036, China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
3.1. Cell lines, cell culture and drugs
3.2. CPE assay of KH901
3.3. Cell viability assay
3.4. Flow cytometry assay
3.5. Western blotting
3.6. Statistical analysis
4. Results
4.1. Synergistic effect of adenovirus KH901 and 5-FU for cell killing
4.2. KH901 induced LoVo cells to enter S-phase
4.3. Expression of p-AKT, Bax, and Bcl-2 was significantly elevated when treated with KH901
5. Discussion
6. Acknowledgements
7. References

1. ABSTRACT

A combination of oncolytic and chemotherapeutic agents has been used to kill cancer cells. However, the effect of oncolytic adenoviruses on the cell cycle remains to be determined. Cytotoxicity assays were performed to determine cell death in cells treated with 5-fluorouracil (5-FU) alone or in combination with the oncolytic adenovirus KH901. Dynamic changes in the cell cycle, cell proliferation, and apoptosis-related proteins including p-AKT, Bcl-2, Bax, and caspase 3 were investigated after treatment with 5-FU with or without KH901. A higher proportion of S-phase cells were observed after treatment with KH901 and 5-FU than with 5-FU alone. p-AKT, Bcl-2, and Bax expression was increased upon treatment with KH901, whereas the expression of caspase-3 was not induced upon treatment with KH901 with or without 5-FU. KH901 exhibited significant potential as an oncolytic adenovirus and increased cell death in combination with 5-FU in LoVo cells, as compared to 5-FU alone. In conclusion, KH901 stimulates LoVo cells to enter the S-phase by activation of p-AKT, which could partly explain its synergistic effect with 5-FU on LoVo cell cytotoxicity.