Laboratory of Endocrinology and Reproductive Biology, Clinical Hospital, University of Chile, Chile.
Gynecol Oncol. 2011 Apr;121(1):13-23. doi: 10.1016/j.ygyno.2010.12.341. Epub 2011 Jan 14.
To evaluate the role of trkA receptor as a potential tumor marker in serous epithelial ovarian cancer and its relationship with the angiogenic factors expression as vascular endothelial growth factor (VEGF) and nerve growth factor (NGF). Additionally, to examine whether NGF and VEGF secreted by epithelial ovarian cancer (EOC) explants and from epithelial ovarian cancer cell line (A2780) are involved in the process of angiogenesis, such as cellular proliferation, migration and differentiation of the human endothelial cell line (EA.hy926).
The mRNA levels of VEGF, NGF and trkA receptors were measured using PCR in 60 ovarian samples. Cellular localization and semi-quantitative estimation of VEGF, NGF, total trkA and p-trkA was performed using IHC in epithelial cells. NGF, total trkA and p-trkA protein were also evaluated in endothelial cells from the same tissues. Human endothelial cell line EA.hy926 was cultured with conditioned media obtained from both EOC explants and from the A2780 cell line, with or without NGF stimulus.
Significantly higher levels of NGF, total trkA and p-trkA protein expressions were observed in epithelial and endothelial cells in poorly differentiated EOC versus normal ovary. Interestingly, the p-trkA receptor expression level showed the most significant difference and its presence was only found in borderline tumor and EOC samples indicating the importance of trkA receptor in EOC as a potential tumor marker. A significant increase in proliferation, migration and differentiation of EA.hy926 cells was observed with NGF, and this effect was significantly reverted when NGF was immuno-blocked and when a trkA inhibitor was used, showing that NGF is an important angiogenic factor in EOC by activating its trkA receptor.
These results indicate that p-trkA may be considered as a new potential tumor marker in EOC, and that NGF may also act as a direct angiogenic factor in EOC.
评估 trkA 受体作为浆液性上皮性卵巢癌潜在肿瘤标志物的作用及其与血管内皮生长因子（VEGF）和神经生长因子（NGF）表达的关系。此外，研究上皮性卵巢癌（EOC）外植体和上皮性卵巢癌细胞系（A2780）分泌的 NGF 和 VEGF 是否参与人内皮细胞系（EA.hy926）的血管生成过程，如细胞增殖、迁移和分化。
使用 PCR 法检测 60 例卵巢样本中 VEGF、NGF 和 trkA 受体的 mRNA 水平。采用免疫组化法检测上皮细胞中 VEGF、NGF、总 trkA 和 p-trkA 的细胞定位和半定量估计。还评估了来自同一组织的内皮细胞中的 NGF、总 trkA 和 p-trkA 蛋白。用 EOC 外植体和 A2780 细胞系获得的条件培养基培养人内皮细胞系 EA.hy926，有或没有 NGF 刺激。
与正常卵巢相比，低分化 EOC 的上皮细胞和内皮细胞中 NGF、总 trkA 和 p-trkA 蛋白表达水平显著升高。有趣的是，p-trkA 受体表达水平差异最显著，仅在交界性肿瘤和 EOC 样本中发现，表明 trkA 受体在 EOC 中作为潜在的肿瘤标志物的重要性。NGF 可显著促进 EA.hy926 细胞的增殖、迁移和分化，当用 NGF 免疫阻断和使用 trkA 抑制剂时，这种作用显著逆转，表明 NGF 通过激活其 trkA 受体是 EOC 中的一种重要血管生成因子。
这些结果表明，p-trkA 可作为 EOC 的一种新的潜在肿瘤标志物，NGF 也可作为 EOC 的直接血管生成因子。
