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HOTAIR/Sp1/miR-199a critically regulates cancer stemness and malignant progression of cutaneous squamous cell carcinoma

Oncogene volume 41, pages 99–111 (2022)Cite this article

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Abstract

The long non-coding RNA (lncRNA), HOX antisense intergenic RNA (HOTAIR) is a well-characterized oncogene in multiple human cancers, but not in cutaneous squamous cell carcinoma (CSCC). In this study, we focused on investigating the potential role of HOTAIR in stemness of CSCC. By measuring its expression using RT-qPCR in CSCC vs. normal tissues, as well as in CSCC cell lines A431 or SCC13, A431- or SCC13-derived CSCC stem cells (CSCSCs), and normal skin fibroblasts (HSFs), we detected higher expression of HOTAIR in CSCC than in normal tissues, in recurrent than in non-recurrent CSCC tissues, in CSCCs and CSCSCs than in HSFs, and particularly, in CSCSCs than in CSCCs. Kaplan–Meier analysis suggested that higher expression of HOTAIR was positively correlated with worse overall survival of CSCC patients. Functional assays on colony formation, EdU incorporation, sphere formation, western blot on stem-cell biomarkers, and in vivo models showed that HOTAIR was essential in maintaining multiple stem cell phenotypes of CSCSCs in vitro and in vivo xenograft growth as well as metastasis. Mechanistically, HOTAIR directly interacted with and up-regulated Sp1. Sp1 then induced DNMT1-mediated promoter methylation and direct transcriptional repression of miR-199a-5p. Targeting Sp1 or DNMT1 further boosted the in vivo anti-tumor and anti-metastasis activities of targeting HOTAIR. In conclusion, HOTAIR, by up-regulating Sp1 and targeting miR-199a, promotes stemness and progression of CSCC. Targeting HOTAIR, Sp1 or the underlying mechanisms may thus benefit CSCC treatment.

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Fig. 1: HOTAIR was up-regulated in CSCC tissues and associated with worse prognosis.
Fig. 2: HOTAIR expression was up-regulated in A431-derived CSCSCs.
Fig. 3: HOTAIR essentially maintained the stem cell features of A431-derived CSCSCs.
Fig. 4: HOTAIR knockdown suppressed xenograft growth and experimental metastasis in vivo.
Fig. 5: HOTAIR directly interacted with Sp1 and up-regulated its expression.
Fig. 6: miR-199a expression was inhibited by DNMT1-mediated promoter methylation.
Fig. 7: Sp1 inhibited miR-199a expression through Sp1-dependent methylation and direct transcriptional repression.
Fig. 8: Co-targeting HOTAIR with Sp1 or DNMT1 inhibited in vivo xenograft growth and metastasis.
Fig. 9: Schematic graph of proposed pathway in CSCC progression.

Data availability

All data generated or analyzed during this study are included in this article. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments. This work was supported by the “perfect medical run-up” talent engineering project of The Third Xiangya Hospital of Central South University (Project Number: JY201719).

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  1. Department of Plastic Surgery, The Third Xiangya Hospital of Central South University, Changsha, 410013, Hunan, PR China

    Jia Chen, Shu-Fen Hou, Feng-Jie Tang, Dai-Song Liu, Zi-Zi Chen, Hong-Lian Zhang & Shao-Hua Wang

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Contributions

Conception and study design: JC; Data acquisition: SFH, FJT; Data analysis: DSL, ZZC; Manuscript drafting: HLZ; Manuscript revising: SHW. All authors have read and approved the final version of this manuscript to be published.

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Correspondence to Shao-Hua Wang.

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This study was reviewed and approved by the Ethics Committee of the Third Xiangya Hospital of Central South University. All patients signed the informed consent to participate in this study. All animal protocols were reviewed and approved by the Institutional Animal Care and Use Committee of Central South University.

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Chen, J., Hou, SF., Tang, FJ. et al. HOTAIR/Sp1/miR-199a critically regulates cancer stemness and malignant progression of cutaneous squamous cell carcinoma. Oncogene 41, 99–111 (2022). https://doi.org/10.1038/s41388-021-02014-x

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