中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (1): 35-37.

• 论著 • 上一篇    下一篇

血管紧张素Ⅱ在A375细胞株的表达及其对血管新生的影响

李东升1,李家文2   

  1. 1. 武汉市第一医院皮肤科
    2. 武汉市华中科技大学同济医学院附属协和医院皮肤科,Affiliated Union Hospital, Tongji Medical College of Huazhong University of Science and Technology
  • 收稿日期:2011-03-14 修回日期:2011-04-14 出版日期:2012-01-15 发布日期:2011-12-31
  • 通讯作者: 李东升 E-mail:bighead18@126.com
  • 基金资助:

    湖北省卫生厅科研基金重点项目

Expression of angiotensinⅡ in A375 human malignant melanoma cells and its influence on angiogenesis

  • Received:2011-03-14 Revised:2011-04-14 Online:2012-01-15 Published:2011-12-31
  • Contact: LI DongSheng E-mail:bighead18@126.com

摘要:

目的 探讨血管紧张素Ⅱ(AngⅡ)在恶性黑素瘤(恶黑)细胞的表达情况及AngⅡ对A375血管新生的影响。 方法 放免法检测人恶黑细胞株A375及人正常黑素细胞培养上清液中AngⅡ的表达水平,体外小管形成实验检测AngⅡ对人脐静脉内皮细胞(HUVEC)体外小管形成的影响,逆转录聚合酶链反应(RT-PCR)及酶联免疫吸附试验(ELISA)法检测A375细胞分别在AngⅡ及AngⅡ受体1(AT1R)特异性拮抗剂洛沙坦作用前后血管内皮细胞生长因子(VEGF)及碱性成纤维细胞生长因子(bFGF)的表达。结果 A375细胞株培养上清液中AngⅡ表达水平为(37.29 ± 0.27) pmol/L,较正常人黑素细胞(21.58 ± 0.32 pmol/L)明显增高,差异有统计学意义(P < 0.05)。AngⅡ能明显诱导HUVEC管状结构的形成,1 μmol/L AngⅡ处理后,二维基质胶中HUVEC形成完整的管状结构,其面积明显高于对照组,为对照组(2.5 ± 0.3)倍,差异有统计学意义(P < 0.05)。AngⅡ处理后,VEGF及bFGF在A375细胞mRNA及蛋白水平的表达较处理前明显增高,1 μmol/L洛沙坦作用后则正好相反,差异有统计学意义(P < 0.05)。结论 A375细胞株存在着AngⅡ的超表达,AngⅡ能明显促进其血管新生,可能是局部肾素-血管紧张素系统影响恶黑肿瘤发生的机制之一。

关键词: 血管

Abstract:

Objective To investigate the expression of angiotensinⅡ (AngⅡ) in malignant melanoma cells and to explore the influence of AngⅡ on angiogenesis. Methods The expression of AngⅡ in the supernatant of A375 cells and primary human melanocytes was detected by radioimmunoassay. Human umbilical vein endothelial cells (HUVECs) were incubated with AngⅡ of 1 ?滋mol/L for 20 hours in an in vitro tube formation assay to observe the effects of AngⅡ on tube formation. A375 cells were incubated with angiotensinⅡ of 1 ?滋mol/L and losartan (an inhibitor of angiotensin II type 1 receptor, AT1R) of 1 ?滋mol/L, respectively for 24 hours; subsequently, reverse transcription PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA)were carried out to measure the mRNA and protein expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), respectively. Results The level of AngⅡ was 37.29 ± 0.27 pmol/L in the supernatant of A375 cells, significantly higher than that in the supernatant of normal human melanocytes (21.58 ± 0.32 pmol/L, P < 0.05). AngⅡ apparently promoted the tube formation by HUVECs. Intact tubiform structures were formed by HUVECs in two-dimension matrigel after being treated with AngⅡ of 1 ?滋mol/L, with the area of tubiform structures being 2.5 ± 0.3 times that in the HUVECs treated with phosphate buffered solution (PBS) (P < 0.05). The protein expressions of VEGF and bFGF in the supernatant of A375 cells and their mRNA expressions in A375 cells were significantly increased by AngⅡ, but suppressed by losartan (all P < 0.05). Conclusions There is a local overexpression of AngⅡ in malignant melanoma, which can markedly promote angiogenesis. This may be one of the mechanisms by which the local renin-angiotensin system affects the initiation of malignant melanoma.