【文献标题】Upregulation of microRNA-141 contributes to suppressing epithelial-mesenchymal transition and lymph node metastasis in laryngeal cancer through HOXC6-dependent TGF-βsignaling pathway

【作者】Li Chen，De-Zhong Sun，Yu-Gui Fu，et.al

【作者单位】临沂市人民医院（Linyi People’s Hospital）

【文献中引用产品】

生物素标记的山羊抗兔

【关键词】microRNA-141, TGF-β signaling pathway, Homeobox C6, epithelial-mesenchymal transition, metastasis, laryngeal cancer

【DOI】doi.org/10.1016/j.cellsig.2019.109444

【影响因子(IF)】6.40

【出版期刊】《Cellular Signalling》

【产品原文引用】

ImmunohistochemistryThe positive expression rates of HOXC6 in laryngeal cancer and adjacent normal tissues were detected by immunohistochemistry. Laryngeal cancer tissues and adjacent normal tissues were fixed in 4% paraformaldehyde for 24 h, dehydrated using 80%, 90% and 100% ethanol successively, cleared with n-butanol, placed in a paraffin box at 60°C and embedded in paraffin. After that, the tissues were cut into 5 μm-thick slices, deparaffinized with xylene after being heated at 60°C for 1 h. The slices were then dehydrated by gradient alcohol and soaked in 3% H2O2 for 10 min, followed by antigen retrieval. Next, the slices were boiled in 0.01 M citric acid buffer (pH = 6.0) at 95°C for 15 - 20 min, and incubated with 100 μL 5% bovine serum albumin (BSA) blocking solution at 37°C for 30 min, followed by incubating with 100 μL primary rabbit antibody to HOXC6 (1 : 50, PB0948, Wuhan Boster Biological Technology, Ltd., Wuhan, Hubei, China) overnight at 4°C. The slices were then incubated with biotin-labeled goat anti-rabbit (1 : 100, HY90046, Shanghai HengYuan Biological Technology Co., Ltd., Shanghai, China) at 37°C for 30 min and with streptavidin-peroxidase solution (Beijing Zhongshan Biological Technology Co., Ltd., Beijing, China) at 37°C for 30 min, and then developed by diaminobenzidine staining solution (Beijing Bioss Biological Technology Co., Ltd., Beijing, China) at room temperature. Subsequently, the slices were soaked in hematoxylin for 5 min, rinsed with 1% hydrochloric acid for 4 s and washed under tap water for 20 min. Brownish-yellow cells were confirmed as HOXC6 positive cells [23]. Five high-magnification visual fields were randomly selected from each slice with 100 cells per field. HOXC6 positive expression was defined by the percentage of positive cells: number of HOXC6 positive tumor cells/number of total tumor cells > 10% was counted as positive (+), < 10% was regarded as negative (-). Experiments were repeated 3 times independently.

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