Basic Research
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 21, 2005; 11(15): 2283-2290
Published online Apr 21, 2005. doi: 10.3748/wjg.v11.i15.2283
Differential gene expression during capillary morphogenesis in a microcarrier-based three-dimensional in vitro model of angiogenesis with focus on chemokines and chemokine receptors
Xi-Tai Sun, Min-Yue Zhang, Chang Shu, Qiang Li, Xiao-Gui Yan, Ni Cheng, Yu-Dong Qiu, Yi-Tao Ding
Xi-Tai Sun, Yi-Tao Ding, Qiang Li, Xiao-Gui Yan, Yu-Dong Qiu, Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Medical College, Hepatobiliary Research Institute, Nanjing University, Nanjing 210008, Jiangsu Province, China
Chang Shu, Ni Cheng, State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing 210009, Jiangsu Province, China
Min-Yue Zhang, State Key Laboratory of Pharmaceutical Biotechnology, Model Animal Research Center, Department of Biochemistry, Nanjing University, Nanjing 210009, Jiangsu Province, China
Author contributions: All authors contributed equally to the work.
Correspondence to: Dr. Yi-Tao Ding, Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Medical College, Hepatobiliary Research Institute, Nanjing University, Nanjing 210008, Jiangsu Province, China. dingyitao@yahoo.com
Telephone: +86-25-83308769 Fax: +86-25-83317016
Received: March 13, 2004
Revised: March 14, 2004
Accepted: April 13, 2004
Published online: April 21, 2005
Abstract

AIM: To globally compare the gene expression profiles during the capillary morphogenesis of human microvascular endothelial cells (HMVECs) in an in vitro angiogenesis system with affymetrix oligonucleotide array.

METHODS: A microcarrier-based in vitro angiogenesis system was developed, in which ECs migrated into the matrix, proliferated, and formed capillary sprouts. The sprouts elongated, branched and formed networks. The total RNA samples from the HMVECs at the selected time points (0.5, 24, and 72 h) during the capillary morphogenesis were used for microarray analyses, and the data were processed with the softwares provided by the manufacturers. The expression patterns of some genes were validated and confirmed by semi-quantitative RT-PCR. The regulated genes were grouped based on their molecular functions and expression patterns, and among them the expression of chemokines and chemokine receptors was specially examined and their functional implications were analyzed.

RESULTS: A total of 1961 genes were up- or downreg-ulated two-folds or above, and among them, 468 genes were up- or down-regulated three-folds or above. The regulated genes could be grouped into categories based on their molecular functions, and were also clustered into six groups based on their patterns of expression. As for chemokines and chemokine receptors, CXCL1/GRO-α, CXCL2/GRO-β, CXCL5/ENA-78, CXCL6/GCP2, IL-8/CXCL8, CXCL12/SDF-1, CXCL9/Mig, CXC11/ITAC, CX3CL1/fractalkine, CCL2/MCP-1, CCL3, CCL5/RANTES, CCL7, CCL15, CCL21, CCL23, CCL28, and CCR1, CCR9, CXCR4 were identified. Moreover, these genes demonstrated different changing patterns during the capillary morphogenesis, which implied that they might have different roles in the sequential process. Among the chemokines identified, CCL2/MCP-1, CCL5/RANTES and CX3CL1 were specially up-regulated at the 24-h time point when the sprouting characterized the morphological change. It was thus suggested that they might exert crucial roles at the early stage of angiogenesis.

CONCLUSION: The present study demonstrates a global profile of gene expression during endothelial capillary morphogenesis, and the results provide us much information about the molecular mechanisms of angiogenesis, with which further evaluation of individual genes can be conducted.

Keywords: Angiogenesis, In vitro model, Endothelial cell, Oligonucleotide array