Wang Chunhui, Chen Lei, Huang Yuanyuan, Li Kun, Jinye Anqi, Fan Taotao, Zhao Ren, Xia Xianming, Shen Bing, Du Juan, Liu Yehai
Department of Otorhinolaryngology, Head and Neck Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China.
Department of Physiology, School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui 230022, P.R. China.
Oncol Lett. 2019 Feb;17(2):1953-1961. doi: 10.3892/ol.2018.9752. Epub 2018 Nov 23.
The prognosis for patients with head and neck cancer (HNC) remains poor, owing to uncontrolled tumor invasion and metastasis. Epithelial-mesenchymal transition (EMT) serves an important role in this invasion and metastasis, and transient receptor potential polycystic 2 (TRPP2) enhances metastasis and invasion by regulating EMT in human laryngeal squamous cell carcinoma. The present study examined whether exosomes/TRPP2 small interfering RNA (siRNA) complexes were able to reduce EMT by suppressing TRPP2 expression in FaDu cells, a cell line of human pharyngeal squamous cell carcinoma. Using agarose gel electrophoresis, it was determined that exosome/TRPP2 siRNA complexes were stable in the presence of nucleases and serum. A fluorescence assay and western blotting analysis was performed, and it was reported that the FaDu cells took up exosomes, the exosomes effectively delivered TRPP2 siRNA into FaDu cells and that exosome/TRPP2 siRNA complexes significantly suppressed TRPP2 protein expression levels in FaDu cells. Furthermore, expression levels of E-cadherin were significantly increased, whereas expression levels of N-cadherin and vimentin were significantly decreased in FaDu cells transfected with TRPP2 siRNA. Thus, exosome/TRPP2 siRNA complexes markedly suppressed TRPP2 expression and EMT in FaDu cells. These results suggested that further development of exosome/TRPP2 siRNA complexes for use as an RNA-based gene therapy in the treatment of HNC is warranted.
由于肿瘤侵袭和转移无法得到控制,头颈癌(HNC)患者的预后仍然很差。上皮-间质转化(EMT)在这种侵袭和转移过程中发挥重要作用,而瞬时受体电位多囊蛋白2(TRPP2)通过调节人喉鳞状细胞癌中的EMT来增强转移和侵袭。本研究检测了外泌体/TRPP2小干扰RNA(siRNA)复合物是否能够通过抑制人咽鳞状细胞癌细胞系FaDu细胞中的TRPP2表达来减少EMT。通过琼脂糖凝胶电泳确定外泌体/TRPP2 siRNA复合物在核酸酶和血清存在的情况下是稳定的。进行了荧光测定和蛋白质印迹分析,结果显示FaDu细胞摄取了外泌体,外泌体有效地将TRPP2 siRNA递送至FaDu细胞中,并且外泌体/TRPP2 siRNA复合物显著抑制了FaDu细胞中TRPP2蛋白的表达水平。此外,在转染了TRPP2 siRNA的FaDu细胞中,E-钙黏蛋白的表达水平显著升高,而N-钙黏蛋白和波形蛋白的表达水平显著降低。因此,外泌体/TRPP2 siRNA复合物显著抑制了FaDu细胞中TRPP2的表达和EMT。这些结果表明,进一步开发外泌体/TRPP2 siRNA复合物作为基于RNA的基因疗法用于治疗头颈癌是有必要的。