Exosome-delivered TRPP2 SiRNA Inhibits Epithelial-mesenchymal Transition Of FaDu Cells

BackgroundAlthough radiotherapy,chemotherapy,immunotherapy,surgery have been achieved and early detection has been improved a lot,the prognosis of head and neck cancer(HNC)was poor due to uncontrolled invasion and metastasis,in which epithelial-mesenchymal transition(EMT)plays an important role.TRPP2 was demonstrated to enhance metastasis and invasion by regulating EMT in human laryngeal squamous cell carcinoma(HLSCC).This finding provides a new idea for the clinical treatment of tumors.ObjectiveTo investigate whether the exosome / TRPP2 si RNA complex can affect the epithelial-mesenchymal transition in Fa Du cells by inhibiting TRPP2 expression.Methods1.Exosomes bind to TRPP2 si RNAExosomes secreted by human embryonic kidney 293 cells(HEK-293 cells)were extracted using exosome extract and high-speed centrifugation and incubated with TRPP2 si RNA.The stability of TRPP2 si RNA was detected by agarose gel electrophoresis.2.Distribution of exosome / TRPP2 si RNA complex in Fa Du cellsFluorescent dye labeled exosomes and si RNA were visualized using a fluorescence microscope to determine if the exosome / TRPP2 si RNA complex was uptake by Fa Du cells.3.To detect the expression of TRPP2,vimentin,N-cadherin and E-cadherin protein in Fa Du cells transfected with exosome / TRPP2 si RNA complexThe expression levels of vimentin,N-cadherin and E-cadherin were detected by Western blotting to determine whether the TPRR2 channel and EMT of transfected Fa Du cells were affected.Results1.Characterization of the exosome/TRPP2 si RNA complex2.Exosomes deliver TRPP2 si RNA into Fa Du cells3.Exosome/TRPP2 si RNA complexes suppress TRPP2 expression and reduce EMT in Fa Du cellsConclusionThe present study demonstrates exosomes/TRPP2 si RNA complex markedly suppresses TRPP2 expression and the EMT process in Fa Du cells.Our results suggest that exosomes/TRPP2 si RNA complex may be used as a RNA-based gene therapy drug in the treatment of head and neck cancer(HNC).