The role of miR-145 in stem cell characteristics of human laryngeal squamous cell carcinoma Hep-2 cells.
The cancer stem-like cells (CSLCs) are tumorigenic cells promoting initiation, progression, and spread of the tumor. Accumulating evidences suggested the presence of CSLCs in distinct tumors including laryngeal squamous cell carcinoma (LSCC). MicroRNAs have been proposed as significant regulators of carcinogenesis, and several of them have been demonstrated to have direct roles in survival of CSLCs. In this study, we aimed to explore the role of miR-145, which is downregulated in LSCC, on cancer stem cell potency of laryngeal cancer cells. We initially showed the downregulation of miR-145 expression in tumor tissue samples and in CD133-enriched CSLCs. Quantitative reverse-transcription PCR (qRT-PCR) analysis of miR-145-transfected Hep-2 cells demonstrated the inhibitory role of miR-145 on stem cell markers like SOX2, OCT4, KLF4, and ABCG2. We, then, investigated the stem cell features of miR-145-overexpressing Hep-2 cells by sphere formation assay, single-cell cloning assay, and aldehyde dehydrogenase (ALDH) assay, which all demonstrated the inhibition of stem cell potency upon miR-145 overexpression. Further qRT-PCR analysis demonstrated altered expression of epithelial to mesenchymal transition markers in miR-145-overexpressing Hep-2 cells. In conclusion, we demonstrated the regulatory role of miR-145 in stem cell characteristics of Hep-2 cells. Based on these results, we propose that miR-145 might carry crucial roles in LSCC tumorigenesis, prognosis, metastasis, chemoresistance, and recurrence through regulating stem cell properties of tumor cells.
Molecular Biology and Genetics Department, Erzurum Technical University, Erzurum, Turkey.
Omer Faruk
OF
Karatas
Department of Medical Genetics, Cerrahpasa Medical School, Istanbul University, Istanbul, Turkey.
Ilknur
I
Suer
Department of Medical Genetics, Cerrahpasa Medical School, Istanbul University, Istanbul, Turkey.|Advanced Genomics and Bioinformatics Research Center, The Scientific and Technological Research Council of Turkey (TUBITAK), Gebze, Kocaeli, Turkey.
Betul
B
Yuceturk
Department of Otorhinolaryngology, Cerrahpasa Medical School, Istanbul University, Istanbul, Turkey.
Mehmet
M
Yilmaz
Department of Otorhinolaryngology, Azerbaijan Medical University, Baku, Azerbaijan.
Yusif
Y
Hajiyev
Department of Medicine and Dan L. Duncan Cancer Center Division of Biostatistics, Baylor College of Medicine, Houston, TX, USA.
Chad J
CJ
Creighton
Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX, 77030, USA.|Michael E. DeBakey VAMC, Houston, TX, 77030, USA.
Michael
M
Ittmann
Department of Medical Genetics, Cerrahpasa Medical School, Istanbul University, Istanbul, Turkey. mozen@bcm.edu.|Department of Pathology and Immunology, Baylor College of Medicine, Houston, TX, 77030, USA. mozen@bcm.edu.|Department of Medical Genetics/Molecular Biology and Genetics, Biruni University, 10. Yil Caddesi Protokol Yolu No: 45, 34010, Topkapi, Istanbul, Turkey. mozen@bcm.edu.
Mustafa
M
Ozen
D000328:Adult; D000368:Aged; D000444:Aldehyde Dehydrogenase / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D015536:Down-Regulation; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D006801:Humans; D000090062:Kruppel-Like Factor 4; D007822:Laryngeal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D035683:MicroRNAs / Q000502:physiology*; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D018874:Spheroids, Cellular / Q000378:metabolism
C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; C541010:MIRN145 microRNA, human; D035683:MicroRNAs; D000444:Aldehyde Dehydrogenase
Cancer stem-like cells; Hsa-miR-145; Laryngeal squamous cell carcinoma; Stem cell markers