Brief Articles
Copyright ©2009 The WJG Press and Baishideng. All rights reserved.
World J Gastroenterol. Jun 14, 2009; 15(22): 2794-2799
Published online Jun 14, 2009. doi: 10.3748/wjg.15.2794
Effect of p27mt gene on apoptosis of the colorectal cancer cell line Lovo
Jun Chen, Wu-Hua Ding, Shao-Yong Xu, Jia-Ning Wang, Yong-Zhang Huang, Chang-Sheng Deng
Jun Chen, Shao-Yong Xu, Department of Gastroenterology, Affiliated People’s Hospital of Yunyang Medical College, Shiyan 442000, Hubei Province, China
Wu-Hua Ding, Obstetrics and Gynecology Department, Affiliated Taihe Hospital of Yunyang Medical College, Shiyan 442000, Hubei Province, China
Jia-Ning Wang, Yong-Zhang Huang, Institute of Clinical Medical Science of Yunyang Medical College, Shiyan 442000, Hubei Province, China
Chang-Sheng Deng, Department of Gastroenterology, Affiliated Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
Author contributions: Chen J designed the research; Chen J, Wang JN, Ding WH, Huang YZ performed the research and analyzed the data; Chen J wrote the paper; Xu SY and Deng CS supervised the whole study.
Correspondence to: Jun Chen, MD, Associate Professor, Associate Chief Physician, Department of Gastroenterology, Affiliated People’s Hospital of Yunyang Medical College, Shiyan 442000, Hubei Province, China. chenjun@medmail.com.cn
Telephone: +86-719-8637529
Fax: +86-719-8666352
Received: April 13, 2009
Revised: May 11, 2009
Accepted: May 18, 2009
Published online: June 14, 2009
Abstract

AIM: To construct p27mt recombinant adenovirus, transfect the colorectal cell line Lovo and observe the effects of p27mt on Lovo cell apoptosis and cell cycle inhibition.

METHODS: We constructed recombinant adenovirus containing p27mt by homologous recombination in bacteria. The colorectal cancer cell line Lovo was infected with recombinant replication-defective adenovirus Ad-p27mt, and expression of p27mt was determined by Western blotting; the inhibitory effect of p27mt on Lovo cells was detected by cytometry. Cell cycle was determined by flow cytometry. DNA fragment analysis identified the occurrence of apoptosis.

RESULTS: The recombinant adenovirus which already contained p27mt target gene was successfully constructed. When multiplicity of infection was ≥ 50, the infection efficiency was 100%. After transfection of Lovo cells with Ad-p27mt the cells had high p27 expression which was identified by immunoblotting assay. PI staining and flow cytometry showed that 77.96% of colorectal cancer cells were inhibited in phase G0/G1, while in the Ad-LacZ group and blank control group, 27.57% and 25.29% cells were inhibited in the same phase, respectively. DNA fragment analysis, flow cytometry and TUNEL assay demonstrated that p27mt is able to induce apoptosis in colorectal cancer cells.

CONCLUSION: p27mt has an obvious blocking effect on colorectal cancer cell cycle, and most cells were inhibited in phase G0/G1. Therefore, p27mt can induce apoptosis in colorectal cells.

Keywords: Apoptosis, Cell cycle, Colorectal cancer, p27mt, Recombinant adenovirus