The radiosensitivity of endothelial cells isolated from human breast cancer and normal tissue in vitro.
We developed a novel method for harvesting endothelial cells from blood vessels of freshly obtained cancer and adjacent normal tissue of human breast, and compared the response of the cancer-derived endothelial cells (CECs) and normal tissue-derived endothelial cells (NECs) to ionizing radiation. In brief, when tissues were embedded in Matrigel and cultured in endothelial cell culture medium (ECM) containing growth factors, endothelial cells grew out of the tissues. The endothelial cells were harvested and cultured as monolayer cells in plates coated with gelatin, and the cells of 2nd-5th passages were used for experiments. Both CECs and NECs expressed almost the same levels of surface markers CD31, CD105 and TEM-8 (tumor endothelial marker-8), which are known to be expressed in angiogenic endothelial cells, i.e., mitotically active endothelial cells. Furthermore, both CECs and NECs were able to migrate into experimental wound in the monolayer culture, and also to form capillary-like tubes on Matrigel-coated plates. However, the radiation-induced suppressions of migration and capillary-like tube formations were greater for CECs than NECs from the same patients. In addition, in vitro clonogenic survival assays demonstrated that CECs were far more radiosensitive than NECs. In summary, we have developed a simple and efficient new method for isolating endothelial cells from cancer and normal tissue, and demonstrated for the first time that endothelial cells of human breast cancer are significantly more radiosensitive than their normal counterparts from the same patients.
Department of Microbiology, Center for Advanced Medical Education by BK21 Project, College of Medicine, Inha University, Incheon 400-712, Republic of Korea.
Moon-Taek
MT
Park
Eun-Taex
ET
Oh
Min-Jeong
MJ
Song
Woo-Jean
WJ
Kim
Young Up
YU
Cho
Sei Joong
SJ
Kim
Jee-Young
JY
Han
Jun-Kyu
JK
Suh
Eun Kyung
EK
Choi
Byung Uk
BU
Lim
Chang Won
CW
Song
Heon Joo
HJ
Park
D015415:Biomarkers / Q000378:metabolism; D001940:Breast / Q000098:blood supply*; D001943:Breast Neoplasms / Q000098:blood supply*; D002465:Cell Movement / Q000528:radiation effects; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000528:radiation effects; D002478:Cells, Cultured; D004307:Dose-Response Relationship, Radiation; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects*; D005260:Female; D006801:Humans; D018919:Neovascularization, Physiologic / Q000528:radiation effects; D011836:Radiation Tolerance; D013997:Time Factors; D046509:Tissue Culture Techniques
D015415:Biomarkers