Stachon A, Schlüter T, Köller M, Weisser H, Krieg M
Institute of Clinical Chemistry, Transfusion, and Laboratory Medicine, University Clinic Bergmannsheil, Ruhr University of Bochum, Bochum, Germany.
Prostate. 2001 Aug 1;48(3):156-64. doi: 10.1002/pros.1094.
Prostate growth seems to be influenced by paracrine factors like IL-6 originating from the microvascular endothelium. Therefore, our efforts were focused on the primary culture and behavior of microvascular endothelial cells (HPEC) derived from tissue of human benign prostatic hyperplasia (BPH). Until now, the isolation and culture of HPEC from BPH have not been reported.
BPH tissue was cut into small cubes and gently squeezed after incubation with dispase. HPEC were cultured from the resulting cell suspension after a stepwise selection by use of superparamagnetic beads coated with antibodies against endothelial specific antigens. HPEC were characterized by flow cytometry and immunohistochemistry. gamma-Glutamyl transpeptidase activity (specific for microvascular endothelium) was measured after dissolution of the HPEC with Triton X-100. After the incubation of HPEC either with ATP, VEGF, or TNF-alpha, the release of IL-6 was measured by enzyme linked immunosorbent assay (ELISA).
HPEC showed a typical endothelial morphology. They were positive for von Willebrand factor, CD31, CD62E (after stimulation with TNF-alpha), alpha-actin and were negative for fibroblastic antigens and PSA. Proliferation was stimulated by vascular endothelial growth factor (VEGF). gamma-Glutamyl transpeptidase activity in HPEC was 6.3 microIU/microg protein, whereas in human umbilical vein endothelial cells (HUVEC) no gamma-glutamyl transpeptidase activity was detectable. The IL-6 secretion of HPEC was stimulated by VEGF and TNF-alpha, but not by ATP and bradykinin.
For the first time, the primary culture of microvascular endothelial cells from BPH tissue was successfully performed. Our results suggest that HPEC may be actively involved in prostate growth, due to the secretion of regulatory factors such as IL-6.
前列腺的生长似乎受到源自微血管内皮的旁分泌因子如白细胞介素 - 6的影响。因此，我们的研究重点是来自人良性前列腺增生（BPH）组织的微血管内皮细胞（HPEC）的原代培养及其行为。到目前为止，尚未见有从BPH组织中分离和培养HPEC的报道。
将BPH组织切成小方块，用 dispase 孵育后轻轻挤压。通过使用包被有抗内皮特异性抗原抗体的超顺磁性珠进行逐步筛选，从所得细胞悬液中培养HPEC。通过流式细胞术和免疫组织化学对HPEC进行鉴定。在用 Triton X - 100溶解HPEC后，测量γ-谷氨酰转肽酶活性（微血管内皮特异性）。在用ATP、血管内皮生长因子（VEGF）或肿瘤坏死因子 - α（TNF - α）孵育HPEC后，通过酶联免疫吸附测定（ELISA）测量白细胞介素 - 6的释放。
HPEC呈现典型的内皮细胞形态。它们对血管性血友病因子、CD31、CD62E（在TNF - α刺激后）、α - 肌动蛋白呈阳性，对成纤维细胞抗原和前列腺特异性抗原（PSA）呈阴性。血管内皮生长因子（VEGF）刺激其增殖。HPEC中的γ - 谷氨酰转肽酶活性为6.3微国际单位/微克蛋白，而在人脐静脉内皮细胞（HUVEC）中未检测到γ - 谷氨酰转肽酶活性。VEGF和TNF - α刺激HPEC分泌白细胞介素 - 6，但ATP和缓激肽则无此作用。
首次成功地从BPH组织中进行了微血管内皮细胞的原代培养。我们的结果表明，由于HPEC分泌如白细胞介素 - 6等调节因子，它们可能积极参与前列腺的生长。
