Bae-Jump V, Segreti E M, Vandermolen D, Kauma S
Department of Obstetrics and Gynecology, Medical College of Virginia Campus at Virginia Commonwealth University, Richmond, Virginia 23298, USA.
Gynecol Oncol. 1999 May;73(2):265-72. doi: 10.1006/gyno.1999.5353.
The overall goal of this study was to investigate the role of the hepatocyte growth factor (HGF)/Met pathway in the pathophysiology of invasive endometrial carcinoma. Our objectives were (1) to examine expression of HGF and Met in surgical endometrial carcinoma specimens and endometrial carcinoma cell lines, and (2) to determine if HGF would stimulate invasion of endometrial carcinoma cell lines in vitro.
Using RT-PCR and Western immunoblotting, endometrial carcinoma specimens and the endometrial carcinoma cell lines KLE, HEC-1A, HEC-1B, and RL-95 were examined for expression of HGF and Met. A Boyden chamber invasion assay using collagen type I coated 8-microm porous membranes was then used to determine if HGF would stimulate cell invasion. Last, we assessed the capacity of endometrial stromal cells, isolated from normal human endometrium, to produce HGF as determined by an enzyme-linked immunosorbent assay and to stimulate invasion of the KLE cell line.
All of the endometrial carcinoma tissue samples were found to express Met mRNA, and two of four samples expressed HGF mRNA. However, the endometrial carcinoma cell lines expressed only Met and not HGF mRNA. Both the endometrial carcinoma tissue specimens and the endometrial carcinoma cell lines expressed the 140-kDa Met protein. HGF induced the invasion of the KLE and HEC-1A cells through the collagen-coated membranes in a dose-dependent fashion. The optimal concentration of HGF was between 10 and 100 ng/ml. HGF (10 ng/ml) stimulated KLE invasion 1.8-fold (P < 0.05) and HEC-1A invasion 6.5-fold (P < 0.05). During exposure to endometrial stromal cell conditioned medium containing HGF as determined by ELISA, invasion of the KLE cell line was stimulated 2.5-fold (P < 0.05).
These results demonstrate that HGF stimulates the invasion of endometrial carcinoma cells in vitro. Since endometrial adenocarcinoma specimens express Met, these findings suggest that the HGF/Met pathway may play a role in the invasive progression of endometrial carcinoma.
本研究的总体目标是探究肝细胞生长因子（HGF）/Met通路在侵袭性子宫内膜癌病理生理学中的作用。我们的目标是：（1）检测手术切除的子宫内膜癌标本和子宫内膜癌细胞系中HGF和Met的表达；（2）确定HGF是否能在体外刺激子宫内膜癌细胞系的侵袭。
采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法，检测子宫内膜癌标本以及子宫内膜癌细胞系KLE、HEC-1A、HEC-1B和RL-95中HGF和Met的表达。然后使用包被有I型胶原的8微米多孔膜的博伊登小室侵袭试验，来确定HGF是否能刺激细胞侵袭。最后，我们通过酶联免疫吸附测定法评估从正常人类子宫内膜分离的子宫内膜基质细胞产生HGF的能力，以及其刺激KLE细胞系侵袭的能力。
所有子宫内膜癌组织样本均检测到Met mRNA表达，4个样本中有2个表达HGF mRNA。然而，子宫内膜癌细胞系仅表达Met，不表达HGF mRNA。子宫内膜癌组织标本和子宫内膜癌细胞系均表达140 kDa的Met蛋白。HGF以剂量依赖方式诱导KLE和HEC-1A细胞穿过胶原包被的膜侵袭。HGF的最佳浓度在10至100 ng/ml之间。HGF（10 ng/ml）刺激KLE细胞侵袭增加1.8倍（P < 0.05），刺激HEC-1A细胞侵袭增加6.5倍（P < 0.05）。通过酶联免疫吸附测定法测定，在暴露于含有HGF的子宫内膜基质细胞条件培养基期间，KLE细胞系的侵袭增加2.5倍（P < 0.05）。
这些结果表明，HGF在体外刺激子宫内膜癌细胞的侵袭。由于子宫内膜腺癌标本表达Met，这些发现提示HGF/Met通路可能在子宫内膜癌的侵袭进展中起作用。
