Down-regulation of Frizzled-7 expression inhibits migration, invasion, and epithelial-mesenchymal transition of cervical cancer cell lines
Down-regulation of Frizzled-7 expression inhibits migration, invasion, and epithelial-mesenchymal transition of cervical cancer cell lines
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DOI：
10.1007/s12032-015-0552-8
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发表时间：
2015-04-01
期刊：
影响因子：
3.4
通讯作者：
Guo, Kejun
中科院分区：
文献类型：
--
作者：
Deng, Boya;Zhang, Siyang;Guo, Kejun
Frizzled-7（FZD 7）是Wnt信号通路的重要受体，参与多种肿瘤的发生和转移。然而，在宫颈癌中发现的信息有限。本研究旨在探讨FZD 7在宫颈癌细胞迁移、侵袭和上皮-间质转化（EMT）中的功能作用。本研究选择FZD 7表达的HeLa和SiHa宫颈癌细胞系。将靶向FZD 7 mRNA的短发夹状RNA（shRNA）转染HeLa和SiHa细胞，经G418筛选获得稳定转染细胞株。进一步进行功能实验以评估FZD 7下调是否影响HeLa和SiHa细胞的迁移、侵袭和EMT。我们的结果表明，FZD 7表达的下调显着抑制细胞侵袭和迁移，并降低两种细胞类型中MMP 2和MMP 9的表达和活性。此外，FZD 7沉默导致间充质标志物包括波形蛋白和Snail的下调，同时增加上皮标志物E-钙粘蛋白的水平。我们进一步发现，减少FZD 7表达抑制JNK和c-jun的磷酸化水平在HeLa和SiHa细胞，如通过蛋白质印迹分析和免疫荧光测定。总之，我们的结果表明，shRNA介导的FZD 7敲低抑制宫颈癌细胞的侵袭，转移和EMT。FZD 7可能为宫颈癌提供一个有希望的治疗靶点。
Frizzled-7 (FZD7) has been demonstrated as a critical receptor of the Wnt signaling and involves in tumorigenesis and metastasis in many cancer types. However, limited information was found in cervical cancer. The aim of this study was to investigate the functional role of FZD7 in migration, invasion, and epithelial-mesenchymal transition (EMT) of cervical cancer cells. HeLa and SiHa cervical carcinoma cell lines with FZD7 expression were chosen in this study. A short hairpin RNA (shRNA) construct targeting FZD7 mRNA was transfected into HeLa and SiHa cells, and the stably transfected cell lines were obtained through G418 screening. Functional experiments were further performed to assess whether FZD7 down-regulation affects the migration, invasion, and EMT of HeLa and SiHa cells. Our results revealed that down-regulation of FZD7 expression significantly inhibited cell invasion and migration, as well as decreased the expression and activities of MMP2 and MMP9 in both cell types. Additionally, FZD7 silencing resulted in down-regulation of mesenchymal markers including Vimentin and Snail while increased the levels of epithelial marker E-cadherin. We further found that decreased FZD7 expression inhibited the phosphorylation levels of JNK and c-jun in both HeLa and SiHa cells, as determined by Western blot analysis and immunofluorescence. Overall, our results indicate that shRNA-mediated knockdown of FZD7 inhibits invasion, metastasis, and EMT of cervical cancer cells. FZD7 may provide a promising therapeutic target in cervical cancer.