title: Nuclear localization signal-enhanced RNA interference of EZH2 and Oct4 in the eradication of head and neck squamous cell carcinoma-derived cancer stem cells.
abstract: Metastasis is the major cause of high mortality in head and neck squamous cell carcinoma (HNSCC), in which HNSCC-derived cancer stem cells (CSCs) may be involved. Several reports have coupled non-viral gene delivery with RNA interference (RNAi) to target specific genes in cancer cells. However, the delivery efficiency of RNAi is limited and remained to be improved. Moreover, the therapeutic effect of non-viral gene delivery approaches on HNSCC-derived CSCs is still uncertain. In this study, we found that EZH2 and Oct4 are upregulated in HNSCC-derived ALDH1+/CD44+ CSC-like cells. Polyurethane-short branch PEI (PU-PEI)-based administration of double-stranded DNA (dsDNA) encoding small interfering RNA (siRNA) against EZH2 and Oct4 (siEZH2/siOct4) led to partial anti-cancer capacity and mild suppression of CSC-like properties. By pre-conjugation of nuclear localization signal (NLS) to siRNA-expressing dsDNA, the anti-cancer efficacy was enhanced due to elevated nuclear delivery. Notably, the NLS-preconjugated siEZH2/siOct4 constructs remarkably repressed epithelial-mesenchymal transdifferentiation (EMT) and radioresistance in ALDH1+/CD44+ CSC-like cells, in which Wnt5A and CyclinD1 may be involved respectively. We furthermore demonstrated that this improved method was capable of reducing tumor growth and metastasis in vivo. Our findings may provide a feasible non-viral gene delivery method to eradicate HNSCC-derived CSCs and improve HNSCC therapy.
authors.affiliation: Department of Dentistry and Institute of Oral Biology, National Yang-Ming University, Taiwan.
authors.forename: Wen-Liang
authors.initials: WL
authors.lastname: Lo
authors.forename: Yueh
authors.initials: Y
authors.lastname: Chien
authors.forename: Guang-Yuh
authors.initials: GY
authors.lastname: Chiou
authors.forename: Ling-Ming
authors.initials: LM
authors.lastname: Tseng
authors.forename: Han-Shui
authors.initials: HS
authors.lastname: Hsu
authors.forename: Yuh-Lih
authors.initials: YL
authors.lastname: Chang
authors.forename: Kai-Hsi
authors.initials: KH
authors.lastname: Lu
authors.forename: Chian-Shiu
authors.initials: CS
authors.lastname: Chien
authors.forename: Mong-Lien
authors.initials: ML
authors.lastname: Wang
authors.forename: Yi-Wei
authors.initials: YW
authors.lastname: Chen
authors.forename: Pin-I
authors.initials: PI
authors.lastname: Huang
authors.forename: Fang-Wei
authors.initials: FW
authors.lastname: Hu
authors.forename: Cheng-Chia
authors.initials: CC
authors.lastname: Yu
authors.forename: Pen-Yuan
authors.initials: PY
authors.lastname: Chu
authors.forename: Shih-Hwa
authors.initials: SH
authors.lastname: Chiou
mesh_terms: D000818:Animals; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D054338:Cell Transdifferentiation / Q000235:genetics; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D000071221:Enhancer of Zeste Homolog 2 Protein; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006258:Head and Neck Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D006801:Humans; D008297:Male; D008422:Materials Testing; D051379:Mice; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology; D019913:Nuclear Localization Signals / Q000235:genetics*; D050814:Octamer Transcription Factor-3 / Q000037:antagonists & inhibitors* / Q000235:genetics*; D063151:Polycomb Repressive Complex 2; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D000077195:Squamous Cell Carcinoma of Head and Neck; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
chemical_list: D004268:DNA-Binding Proteins; D019913:Nuclear Localization Signals; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D034741:RNA, Small Interfering; D014157:Transcription Factors; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D063151:Polycomb Repressive Complex 2