Interaction between stromal fibroblasts and colorectal cancer cells in the expression of vascular endothelial growth factor.
BACKGROUND
Vascular endothelial growth factor (VEGF), a potent angiogenic factor, has been implicated in metastasis of colorectal cancer (CRC). The present study aimed to clarify whether cancer-stromal interaction induces the production of VEGF.


METHODS
Human colonic fibroblasts (CCD-18Co) and CRC (SW480, SW620) cells were analyzed in this study. The cell cycle of colonic fibroblasts during co-culture was analyzed by flow cytometry. VEGF and TGF-beta1 released into the conditioned media in co-culture models were measured. Northern blot with human specific VEGF probe was performed to identify the expression of VEGF in this model.


RESULTS
Co-culture of colonic fibroblasts with CRC cells increased the viability of fibroblasts during co-culture. Cell cycle analysis revealed that most of the fibroblasts co-cultured with CRC cells were arrested at G1 phase and few cells were in sub-G1 phase that indicates apoptosis. Although VEGF protein was detected in the culture media of all of the monocultures, co-cultivation of CRC with fibroblasts resulted in synergistic increase of VEGF production compared with monocultures. However TGF-beta1 protein was not detected in any conditioned medium. VEGF mRNA was detected in both CRC and fibroblasts. Under co-culture condition, an abundance of VEGF mRNA expression was noted in fibroblasts relative to CRC cells.


CONCLUSIONS
The present study suggests that CRC manipulates the host stroma to suppress apoptosis and up-regulate VEGF production.
Koshida
Yoshitomo
Y
Department of Surgery, Kitasato University School of Medicine, Kanagawa, Japan.
Kuranami
Masaru
M
Watanabe
Masahiko
M
D017209:Apoptosis; D015152:Blotting, Northern; D002453:Cell Cycle; D002460:Cell Line; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D003106:Colon / Q000166:cytology; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology*; D017077:Culture Media, Conditioned; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D005434:Flow Cytometry; D016193:G1 Phase; D015870:Gene Expression; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis; D017154:Stromal Cells / Q000378:metabolism / Q000473:pathology; D016212:Transforming Growth Factor beta / Q000378:metabolism; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
D017077:Culture Media, Conditioned; D012333:RNA, Messenger; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A