Arabzadeh Somayeh, Hossein Ghamartaj, Salehi-Dulabi Zahra, Zarnani Amir Hassan
Department of Animal Physiology, Developmental Biology Laboratory, School of Biology, College of Science, University of Tehran, Tehran, Iran.
Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Cell Mol Biol Lett. 2016 Jul 28;21:9. doi: 10.1186/s11658-016-0003-3. eCollection 2016.
Wnt5A, which is a member of the non-transforming Wnt protein family, is implicated in inflammatory processes. It is also highly expressed by ovarian cancer cells. ROR2, which is a member of the Ror-family of receptor tyrosine kinases, acts as a receptor or co-receptor for Wnt5A. The Wnt5A-ROR2 signaling pathway plays essential roles in the migration and invasion of several types of tumor cell and influences their cell polarity. We investigated the modulation of Wnt5A-ROR2 by inflammatory mediators and its involvement in the migration of the human ovarian cancer cell line SKOV-3.
SKOV-3 cells were treated with LPS (lipopolysaccharide), LTA (lipoteichoic acid) and recombinant human IL-6 alone or in combination with STAT3 inhibitor (S1155S31-201) or NF-kB inhibitor (BAY11-7082) for 4, 8, 12, 24 and 48 h. The Wnt5A and ROR2 expression levels were determined at the gene and protein levels. Cells were transfected with specific siRNA against Wnt5A in the absence or presence of human anti-ROR2 antibody and cell migration was assessed using transwells.
There was a strong downregulation of Wnt5A expression in the presence of STAT3 or NF-kB inhibitors. Cell stimulation with LTA or IL-6 for 8 h led to significantly increased levels of Wnt5A (5- and 3-fold higher, respectively). LPS, LTA or IL-6 treatment significantly increased ROR2 expression (2-fold after 48 h). LPS- or LTA-induced Wnt5A or ROR2 expression was abrogated in the presence of STAT3 inhibitor ( < 0.001). IL-6-induced Wnt5A expression was abrogated by both STAT3 and NF-kB inhibitors ( < 0.001). Although not significant, IL-6-induced ROR2 expression showed a modest decrease when STAT3 inhibitor was used. Moreover, cell migration was decreased by 80 % in siRNA Wnt5A-transfected cells in the presence of anti-human ROR2 antibody ( < 0.001).
This study revealed for the first time that inflammatory mediators modulate Wnt5A and ROR2 through NF-kB and STAT3 transcription factors and this may play a role in ovarian cancer cell migration. The results described here provide new insight into the role of the Wnt5A-ROR2 complex in ovarian cancer progression in relation to inflammation.
Wnt5A是一种非转化型Wnt蛋白家族成员，与炎症过程有关。它在卵巢癌细胞中也高度表达。ROR2是受体酪氨酸激酶Ror家族的成员，作为Wnt5A的受体或共受体发挥作用。Wnt5A - ROR2信号通路在几种类型肿瘤细胞的迁移和侵袭中起重要作用，并影响其细胞极性。我们研究了炎症介质对Wnt5A - ROR2的调节作用及其在人卵巢癌细胞系SKOV - 3迁移中的作用。
将SKOV - 3细胞单独或与STAT3抑制剂（S1155S31 - 201）或NF - kB抑制剂（BAY11 - 7082）联合用脂多糖（LPS）、脂磷壁酸（LTA）和重组人IL - 6处理4、8、12、24和48小时。在基因和蛋白水平测定Wnt5A和ROR2的表达水平。在不存在或存在人抗ROR2抗体的情况下，用针对Wnt5A的特异性siRNA转染细胞，并使用Transwell小室评估细胞迁移。
在存在STAT3或NF - kB抑制剂的情况下，Wnt5A表达强烈下调。用LTA或IL - 6刺激细胞8小时导致Wnt5A水平显著升高（分别高出5倍和3倍）。LPS、LTA或IL - 6处理显著增加ROR2表达（48小时后增加2倍）。在存在STAT3抑制剂的情况下，LPS或LTA诱导的Wnt5A或ROR2表达被消除（<0.001）。IL - 6诱导的Wnt5A表达被STAT3和NF - kB抑制剂均消除（<0.001）。虽然不显著，但当使用STAT3抑制剂时，IL - 6诱导的ROR2表达有适度下降。此外，在存在抗人ROR2抗体的情况下，siRNA Wnt5A转染细胞的迁移减少了80%（<0.001）。
本研究首次揭示炎症介质通过NF - kB和STAT3转录因子调节Wnt5A和ROR2，这可能在卵巢癌细胞迁移中起作用。此处描述的结果为Wnt5A - ROR2复合物在与炎症相关的卵巢癌进展中的作用提供了新的见解。
Zotero 插件
