A proteomic study on cell cycle progression of endothelium exposed to tumor conditioned medium and the possible role of cyclin D1/E.
This study was designed to comprehensively analyze the differential expression of proteins from human umbilical vein endothelial cells (HUVECs) exposed to tumor conditioned medium (TCM) and to identify the key regulator in the cell cycle progression. The HUVECs were exposed to TCM from breast carcinoma cell line MDA-MB-231, then their cell cycle distribution was measured by flow cytometer (FCM). The role of protein in cell cycle progression was detected via two-dimensional polyacrylamide gel electrophoresis (2-DE) and western blotting. Following the stimulation of TCM, HUVECs showed a more cells in the S phase than did the negative control group (ECGF-free medium with 20% FBS), but the HUVECs' level was similar to the positive control group (medium with 25 micrograms/ml ECGF and 20% FBS). Increased expression of cyclin D1/E and some changes in other related proteins occurred after incubation with TCM. From our results, we can conclude that breast carcinoma cell line MDA-MB-231 may secrete soluble pro-angiogenic factors that induce the HUVEC angiogenic switch, during which the expression of cell cycle regulator cyclin D1/E increases and related proteins play an important role in this process.
Li
Ailing
A
Institute of Microcirculation, Chinese Academy Medical Sciences (CAMS) and Peking Union Medical College (PUMC), Beijing 100005, China.
Li
Hongwei
H
Jin
Gang
G
Xiu
Ruijuan
R
D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000473:pathology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor / Q000378:metabolism; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D019938:Cyclin D1 / Q000096:biosynthesis / Q000235:genetics; D019927:Cyclin E / Q000096:biosynthesis / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects*; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D040901:Proteomics; D013382:Subtraction Technique; D014471:Umbilical Veins
D043925:Angiogenesis Inducing Agents; D017077:Culture Media, Conditioned; D019927:Cyclin E; D011506:Proteins; D019938:Cyclin D1; D006861:Hydrogen Peroxide