Department of Gynecology and Obstetrics, Faculty of Medicine, Kyoto University, Kyoto, Japan.
Int J Gynecol Cancer. 2013 Mar;23(3):576-82. doi: 10.1097/IGC.0b013e3182849e36.
The Eph-ephrin system is a unique system that can induce multiple cellular responses such as cell migration, regulation of angiogenesis, and axonal guidance. Previously, the Eph-ephrin system was reported to regulate human extravillous trophoblast invasion. In this study, we examined the possible involvement of the Eph-ephrin system in the invasion of malignant gestational trophoblastic diseases using a human choriocarcinoma-derived cell line, JEG-3.
The mRNA expression of class A Ephs and ephrins on JEG-3 cells was examined by reverse transcription-polymerase chain reaction. The effects of recombinant human Eph A1 (r-Eph A1) and r-ephrin A4 on the proliferation and invasion of JEG-3 cells were investigated by cell proliferation and Matrigel invasion assays. The alterations of integrin expression on JEG-3 cells in the presence of r-Eph A1 and r-ephrin A4 were investigated by flow cytometry. The induction of phosphorylation of focal adhesion kinase in JEG-3 cells by r-ephrin A4 was examined by Western blot analysis.
By reverse transcription-polymerase chain reaction, mRNAs of Eph A1, A2, and A4 and ephrin A1, A4, and A5 were detected on JEG-3 cells. In Matrigel invasion assay, both r-Eph A1 and r-ephrin A4 promoted the invasion of JEG-3 cells without affecting cell proliferation. During 24-hour culture with r-Eph A1 and r-ephrin A4, the increase in integrin α 5 expression on JEG-3 cells was observed by flow cytometry. Western blotting analysis showed that r-ephrin A4 induced dephosphorylation of focal adhesion kinase in JEG-3 cells.
These findings suggest that Eph-ephrin interaction plays some role in the regulation of choriocarcinoma invasion in cooperation with integrins.
Eph-ephrin 系统是一个独特的系统，可以诱导多种细胞反应，如细胞迁移、血管生成的调节和轴突导向。先前，Eph-ephrin 系统被报道调节人绒毛外滋养细胞的侵袭。在这项研究中，我们使用人绒毛膜癌细胞系 JEG-3 检查 Eph-ephrin 系统在恶性妊娠滋养细胞疾病侵袭中的可能作用。
通过逆转录-聚合酶链反应检查 JEG-3 细胞中 EphA 类和 Ephrin 的 mRNA 表达。通过细胞增殖和 Matrigel 侵袭试验研究重组人 EphA1(r-EphA1)和 r-ephrin A4 对 JEG-3 细胞增殖和侵袭的影响。通过流式细胞术研究 r-EphA1 和 r-ephrin A4 存在时 JEG-3 细胞整合素表达的变化。通过 Western blot 分析研究 r-ephrin A4 诱导 JEG-3 细胞中粘着斑激酶的磷酸化。
通过逆转录-聚合酶链反应，在 JEG-3 细胞中检测到 EphA1、A2 和 A4 以及 Ephrin A1、A4 和 A5 的 mRNA。在 Matrigel 侵袭试验中，r-EphA1 和 r-ephrin A4 均促进 JEG-3 细胞的侵袭，而不影响细胞增殖。在与 r-EphA1 和 r-ephrin A4 孵育 24 小时期间，通过流式细胞术观察到 JEG-3 细胞中整合素α5表达增加。Western blot 分析表明，r-ephrin A4 诱导 JEG-3 细胞中粘着斑激酶的去磷酸化。
这些发现表明 Eph-ephrin 相互作用在与整合素协同调节绒毛膜癌侵袭中起一定作用。
