• 重慶醫(yī)科大學附屬第一醫(yī)院肝膽外科( 重慶 400016);

目的 　檢測腫瘤細胞上清液對成纖維細胞的激活情況及激活后血管內(nèi)皮生長因子-A（VEGF-A）表達的變化規(guī)律。
方法 　MTT法檢測普通培養(yǎng)液、含轉(zhuǎn)化生長因子-β1（TGF-β1）的普通培養(yǎng)液以及腫瘤細胞上清液組成的條件培養(yǎng)液對成纖維細胞生長情況的影響，用RT-PCR、Western blot及免疫組織化學法檢測不同培養(yǎng)條件下成纖維細胞的α-平滑肌肌動蛋白（α-SMA）與VEGF-A的表達。
結(jié)果 　腫瘤細胞上清液對成纖維細胞的生長有一定的促進作用。含TGF-β1的普通培養(yǎng)液比普通培養(yǎng)液更有利于成纖維細胞轉(zhuǎn)化為肌成纖維細胞，并且能維持肌成纖維細胞的表型，但是二者均不表達VEGF-A；條件培養(yǎng)液能促進成纖維細胞穩(wěn)定表達α-SMA和VEGF-A，二者在培養(yǎng)后第1天均開始表達，第3天表達量達到峰值，第3天以后表達穩(wěn)定。
結(jié)論 　腫瘤細胞上清液能夠有效、穩(wěn)定地激活成纖維細胞為肌成纖維細胞，成纖維細胞的激活程度影響VEGF-A的表達，二者均具有最佳的激活時間點，并且最佳時間點相一致，最佳激活點的成纖維細胞有望成為一種可用于移植的促進血管重建的細胞。

引用本文： 肖衡,杜成友,羅詩樵,劉定志,潘龍. 腫瘤細胞上清液對成纖維細胞表型轉(zhuǎn)變及血管內(nèi)皮生長因子-A 表達的影響. 中國普外基礎(chǔ)與臨床雜志, 2012, 19(2): 129-134. doi: 復制

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.
  1. 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. 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. 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. 4. Xouri G, Christian S. Origin and function of tumor stroma fibroblasts[J]. Semin Cell Dev Biol, 2010, 21(1)：40-46.
  5. 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. 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. 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. 8. Turner N, Grose R. Fibroblast growth factor signalling：from development to cancer[J]. Nat Rev Cancer, 2010, 10(2)：116-129.
  9. 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. 10. Bhowmick NA, Neilson EG, Moses HL. Stromal fibroblasts in cancer initiation and progression[J]. Nature, 2004, 432(7015)： 332-337.
  11. 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. 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. 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. 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.