Gastric Cancer
Copyright ©The Author(s) 2003. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Nov 15, 2003; 9(11): 2419-2423
Published online Nov 15, 2003. doi: 10.3748/wjg.v9.i11.2419
Construction and analysis of SSH cDNA library of human vascular endothelial cells related to gastrocarcinoma
Yong-Bo Liu, Zhao-Xia Wei, Li Li, Hang-Sheng Li, Hui Chen, Xiao-Wen Li
Yong-Bo Liu, Hui Chen, Xiao-Wen Li, Department of Cell Biology and Medical Genetics, Medical College of Zhengzhou University, Zhengzhou, 450052, Henan Province, China
Zhao-Xia Wei, Department of Anatomy, Henan Medical College for Enterprise Employees, Zhengzhou 450003, Henan Province, China
Li Li, Department of Cardiovascular Disease, People Hospital of Henan Province, Zhengzhou 450003, Henan Province, China
Hang-Sheng Li, Department of Gynecology and Obstetrics, Henan Medical College for Enterprise Employees, Zhengzhou 450003, Henan Province, China
Author contributions: All authors contributed equally to the work.
Correspondence to: Yong-Bo Liu, Department of Cell Biology and Medical Genetics, Medical College of Zhengzhou University, Zhengzhou, 450052, Henan Province, China. liuyb415@163.net
Telephone: +86-371-6974324
Received: May 11, 2003
Revised: May 27, 2003
Accepted: June 4, 2003
Published online: November 15, 2003
Abstract

AIM: To construct subtracted cDNA libraries of human vascular endothelial cells (VECs) related to gastrocarcinoma using suppression substractive hybridization (SSH) and to analyze cDNA libraries of gastrocarcinoma and VECs in Cancer Gene Anatomy Project (CGAP) database.

METHODS: Human VECs related to gastric adenocarcinoma and corresponding normal tissue were separated by magnetic beads coupled with antibody CD31 (Dynabeads CD31). A few amount of total RNA were synthesized and amplified by SMARTTM PCR cDNA Synthesis Kit. Then, using SSH and T/A cloning techniques, cDNA fragments of differentially expressed genes in human VECs of gastric adenocarcinoma were inserted into JM109 bacteria. One hundred positive bacteria clones were randomly picked and identified by colony PCR method. To analyze cDNA libraries of gastrocarcinoma and VECs in CGAP database, the tools of Library Finder, cDNA xProfiler, Digital GENE Expression Displayer (DGED), and Digital Differential Display (DDD) were used.

RESULTS: Forward and reverse subtraction cDNA libraries of human VECs related to gastrocarcinoma were constructed successfully with SSH and T/A cloning techniques. Analysis of CGAP database indicated that no appropriate library of VECs related to carcinoma was constructed.

CONCLUSION: Construction of subtraction cDNA libraries of human VECs related to gastrocarcinoma was successful and necessary, which laid a foundation for screening and cloning new and specific genes of VECs related to gastrocarcinoma.

Keywords: $[Keywords]