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Cell Physiol Biochem. 2018;47(4):1696-1710. doi: 10.1159/000490992. Epub 2018 Jun 27.

Whole-Transcriptome Analysis of CD133+CD144+ Cancer Stem Cells Derived from Human Laryngeal Squamous Cell Carcinoma Cells.

Wu Y1,2,3,4, Zhang Y1,2,3,4, Niu M1,2,3,4, Shi Y1,2,3,4, Liu H1,2,3,4, Yang D1,2,3,4, Li F1,2,3,4, Lu Y5, Bo Y6, Zhang R4,7, Li Z4,8, Luo H1,2,3,4, Cui J1,2,3,4, Sang J1,2,3,4, Xiang C1,2,3,4, Gao W1,2,3,4, Wen S1,2,3,4.

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Shanxi Key Laboratory of Otorhinolaryngology Head and Neck Cancer, Taiyuan, China.
Department of Otolaryngology Head & Neck Surgery, The First Hospital, Shanxi Medical University, Taiyuan, China.
Otolaryngology Head & Neck Surgery Research Institute, Shanxi Medical University, Taiyuan, China.
The Key Scientific and Technological Innovation Platform for Precision Diagnosis and Treatment of Head and Neck Cancer, Shanxi Province, Taiyuan, China.
Department of Otolaryngology Head & Neck Surgery, The First Hospital, Jinzhou Medical University, Jinzhou, China.
Department of Pathology, Shanxi Cancer Hospital, Shanxi Medical University, Taiyuan, China.
Department of MRI & CT, Shanxi Cancer Hospital, Shanxi Medical University, Taiyuan, China.
Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan, China.



CD133+CD44+ cancer stem cells previously isolated from laryngeal squamous cell carcinoma (LSCC) cell lines showed strong malignancy and tumorigenicity. However, the molecular mechanism underlying the enhanced malignancy remained unclear.


Cell proliferation assay, spheroid-formation experiment, RNA sequencing (RNA-seq), miRNA-seq, bioinformatic analysis, quantitative real-time PCR, migration assay, invasion assay, and luciferase reporter assay were used to identify differentially expressed mRNAs, lncRNAs, circRNAs and miRNAs, construct transcription regulatory network, and investigate functional roles and mechanism of circRNA in CD133+CD44+ laryngeal cancer stem cells.


Differentially expressed genes in TDP cells were mainly enriched in the biological processes of cell differentiation, regulation of autophagy, negative regulation of cell death, regulation of cell growth, response to hypoxia, telomere maintenance, cellular response to gamma radiation, and regulation of apoptotic signaling, which are closely related to the malignant features of tumor cells. We constructed the regulatory network of differentially expressed circRNAs, miRNAs and mRNAs. qPCR findings for the expression of key genes in the network were consistent with the sequencing data. Moreover, our data revealed that circRNA hg19_circ_0005033 promotes proliferation, migration, invasion, and chemotherapy resistance of laryngeal cancer stem cells.


This study provides potential biomarkers and targets for LSCC diagnosis and therapy, and provide important evidences for the heterogeneity of LSCC cells at the transcription level.


CD133+CD44+; Cancer stem cells; Circular RNA; Laryngeal squamous cell carcinoma; Whole transcriptome analysis

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