Influences of Tumor Cells Supernatant on Phenotypic Transition of Fibroblasts and Expression of Vascular Endothelial Growth FactorA_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

XIAO Heng , DU Chengyou , LUO Shiqiao , LIU Dingzhi , PAN Long

Department of Hepatobiliary Surgery，The First Affiliated Hospital of Chongqing Medical University，Chongqing 400016，China;

Corresponding author：

Keywords：

Fibroblast; Myofibroblast; Transforming growth factor-β1; Vascular endothelial growth factor-A; α-smooth muscle actin

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Objective 　To investigate the expression of vascular endothelial growth factor-A （VEGF-A） and the phenotypic transition after the activation of fibroblasts by the supernatant of cultured tumor cells．
Methods 　The growth tendency of fibroblasts was tested by the MTT assay．The expressions of alpha-smooth muscle actin （α-SMA） and VEGF-A mRNA were tested by RT-PCR．The expressions of α-SMA and VEGF-A protein were tested by immunohistochemistry and Western blot．
Results 　The MTT assay indicated that the conditional medium which contained tumor cells supernatant could obviously promote the growth of the fibroblasts． RT-PCR and Western blot manifested that α-SMA expressed by the fibroblasts which cultured by normal medium reached its peak on day 5，then decreased to a low level on day 7．When the medium contained 2 ng/ml transforming growth factor-β1 （TGF-β1），the fibroblasts could steadily express more α-SMA．But the above two mediums could not make the fibroblasts express the VEGF-A. When using the conditional medium，the α-SMA peak advanced on the third day and maintained at a high level，so as the expression of the VEGF-A．
Conclusions 　The results suggested that fibroblasts can be activated to be myofibroblasts when using the conditional medium．The best activation time of the fibroblasts is consistent with the time of the VEGF-A expression at the highest level by the activated fibroblasts．The fibroblasts which activated at the best time are expected to become a kind of cells which can be used for promoting revascularization．

Citation： XIAO Heng,DU Chengyou,LUO Shiqiao,LIU Dingzhi,PAN Long,.. Influences of Tumor Cells Supernatant on Phenotypic Transition of Fibroblasts and Expression of Vascular Endothelial Growth FactorA. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2012, 19(2): 129-134. doi: Copy

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1. Meyer-Ter-Vehn T, Gebhardt S, Sebald W, et al. p38 inhibitors prevent TGF-β-induced myofibroblast transdifferentiation in human tenon fibroblasts[J]. Invest Ophthalmol Vis Sci, 2006, 47(4)：1500-1509.
2. Ishida Y, Gao JL, Murphy PM. Chemokine receptor CX3CR1 mediates skin woundhealingby promoting macrophageand fibroblast accumulation and function[J]. J Immunol, 2008, 180(1)：569-579.
3. Viacava P, Naccarato AG, Bocci G, et al. Angiogenesis and VEGF expression in pre-invasive lesions of the human breast[J]. J Pathol, 2004, 204(2)：140-146.
4. Xouri G, Christian S. Origin and function of tumor stroma fibroblasts[J]. Semin Cell Dev Biol, 2010, 21(1)：40-46.
5. Shao J, Sheng GG, Mifflin RC, et al. Roles of myofibroblasts in prostaglandin E2-stimulated intestinal epithelial proliferation and angiogenesis[J]. Cancer Res, 2006, 66(2)：846-855.
6. Silzle T, Randolph GJ, Kreutz M, et al. The fibroblast：sentinel cell and local immune modulator in tumor tissue[J]. Int J Cancer, 2004, 108(2)：173-180.
7. Fukumura D, Xavier R, Sugiura T, et al. Tumor induction of VEGF promoter activity in stromal cells[J]. Cell, 1998, 94(6)： 715-725.
8. Turner N, Grose R. Fibroblast growth factor signalling：from development to cancer[J]. Nat Rev Cancer, 2010, 10(2)：116-129.
9. Rizzino A, Kazakoff P, Ruff E, et al. Regulatory effects of cell density on the binding of transforming growth factor β, epidermal growth factor, platelet-derived growth factor, and fibroblast growth factor[J]. Cancer Res, 1988, 48(15)：4266-4271.
10. Bhowmick NA, Neilson EG, Moses HL. Stromal fibroblasts in cancer initiation and progression[J]. Nature, 2004, 432(7015)： 332-337.
11. Masur KS, Dewal HS, Dinh TT, et al. Myofibroblasts differentiate from fibroblasts when plated at low density[J]. Proc Natl Acad Sci USA, 1996, 93(9)：4219-4223.
12. Sonoshita M, Takaku K, Sasaki N, et al. Acceleration of intestinal polyposis through prostaglandin receptor EP2 in ApcΔ716 knockout mice[J]. Nat Med, 2001, 7(9)：1048-1051.
13. Seno H, Oshima M, IshikawaTO, et al. Cyclooxygenase 2- and prostaglandin E2 receptor EP2-dependent angiogenesis in ApcΔ716 mouse intestinal polyps[J]. Cancer Res, 2002, 62：506.
14. Puca R, Nardinocchi L, D’Orazi G. Regulation of vascular endothelial growth factor expression by homeodomain-interacting protein kinase-2[J]. J Exp Clin Cancer Res, 2008, 27：22.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Influences of Tumor Cells Supernatant on Phenotypic Transition of Fibroblasts and Expression of Vascular Endothelial Growth FactorA

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