Zheng LD, Jiang GS, Pu JR, Mei H, Dong JH, Hou XH, Tong QS. Stable knockdown of heparanase expression in gastric cancer cells in vitro. World J Gastroenterol 2009; 15(43): 5442-5448 [PMID: 19916174 DOI: 10.3748/wjg.15.5442]
Corresponding Author of This Article
Dr. Qiang-Song Tong, Department of Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China. qs_tong@hotmail.com
Article-Type of This Article
Original Article
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Li-Duan Zheng, Department of Pathology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Guo-Song Jiang, Jia-Rui Pu, Hong Mei, Qiang-Song Tong, Department of Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Ji-Hua Dong, Department of Central Laboratory, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Xiao-Hua Hou, Department of Gastroenterology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Author contributions: Tong QS designed the study and drafted the manuscript; Zheng LD, Jiang GS, Pu JR and Mei H performed the research; Dong JH and Hou XH engaged in drafting the manuscript and in statistical analysis; Zheng LD and Jiang GS contributed equally to this work.
Supported by The National Natural Science Foundation of China, No. 30200284, No. 30600278, No. 30772359, Program for New Century Excellent Talents in University, NCET-06-0641, and Scientific Research Foundation for the Returned Overseas Chinese Scholars, 2008-889
Correspondence to: Dr. Qiang-Song Tong, Department of Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China. qs_tong@hotmail.com
Telephone: +86-27-63776478 Fax: +86-27-85726197
Received: August 30, 2009 Revised: October 15, 2009 Accepted: October 21, 2009 Published online: November 21, 2009
Abstract
AIM: To develop short hairpin RNA (shRNA) against heparanase, and to determine its effects on heparanase expression and the malignant characteristics of gastric cancer cells.
METHODS: Heparanase-specific shRNA was constructed and transferred into cultured the gastric cancer cell line SGC-7901. Stable subclonal cells were screened by G418 selection. Heparanase expression was measured by reverse transcriptase-polymerase chain reaction (RT-PCR), real-time quantitative PCR and Western blotting. Cell proliferation was detected by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetry and colony formation assay. The in vitro invasiveness and metastasis of cancer cells were measured by cell adhesion assay, wound healing assay and matrigel invasion assay. The angiogenesis capabilities of cancer cells were measured by tube formation of endothelial cells.
RESULTS: Stable transfection of heparanase-specific shRNA, but not of scrambled shRNA and mock vector, resulted in reduced mRNA and protein levels of heparanase. The shRNA-mediated knockdown of heparanase did not affect the cellular proliferation of SGC-7901 cells. However, the in vitro invasiveness and metastasis of cancer cells were decreased after knockdown of heparanase. Moreover, transfection of heparanase-specific shRNA decreased the in vitro angiogenesis capabilities of SGC-7901 cells.
CONCLUSION: Stable knockdown of heparanase can efficiently decrease the invasiveness, metastasis and angiogenesis of human gastric cancer cells. In contrast, stable knockdown of heparanase does not affect the cell proliferation.
