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卷曲蛋白-7(Frizzled-7)表达下调可抑制宫颈癌细胞系的迁移、侵袭及上皮-间质转化。

Down-regulation of Frizzled-7 expression inhibits migration, invasion, and epithelial-mesenchymal transition of cervical cancer cell lines.

作者信息

Deng Boya, Zhang Siyang, Miao Yuan, Zhang Yi, Wen Fang, Guo Kejun

机构信息

Department of Gynecology, The First Affiliated Hospital of China Medical University, 155 North Nanjing Street, Shenyang, 110001, People's Republic of China,

出版信息

Med Oncol. 2015 Apr;32(4):102. doi: 10.1007/s12032-015-0552-8. Epub 2015 Mar 5.

Abstract

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.

摘要

卷曲蛋白7(FZD7)已被证明是Wnt信号通路的关键受体,并参与多种癌症类型的肿瘤发生和转移。然而,关于宫颈癌的相关信息有限。本研究的目的是探讨FZD7在宫颈癌细胞迁移、侵袭和上皮-间质转化(EMT)中的功能作用。本研究选用了具有FZD7表达的HeLa和SiHa宫颈癌细胞系。将靶向FZD7 mRNA的短发夹RNA(shRNA)构建体转染到HeLa和SiHa细胞中,并通过G418筛选获得稳定转染的细胞系。进一步进行功能实验,以评估FZD7下调是否会影响HeLa和SiHa细胞的迁移、侵袭和EMT。我们的结果显示,FZD7表达下调显著抑制了细胞侵袭和迁移,同时降低了两种细胞类型中MMP2和MMP9的表达及活性。此外,FZD7沉默导致间充质标志物波形蛋白和蜗牛蛋白表达下调,而上皮标志物E-钙黏蛋白水平升高。通过蛋白质印迹分析和免疫荧光检测,我们进一步发现,FZD7表达降低抑制了HeLa和SiHa细胞中JNK和c-jun的磷酸化水平。总体而言,我们的结果表明,shRNA介导的FZD7敲低抑制了宫颈癌细胞的侵袭、转移和EMT。FZD7可能为宫颈癌提供一个有前景的治疗靶点。

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