GLI3 knockdown decreases stemness, cell proliferation and invasion in oral squamous cell carcinoma

Author(s)

Setubal Destro Rodrigues, Maria Fernanda ^{[1, 2]} ; Miguita, Lucyene ^[1] ; De Andrade, Nathalia Paiva ^[1] ; Heguedusch, Daniele ^[1] ; Rodini, Camila Oliveira ^[3] ; Moyses, Raquel Ajub ^[4] ; Toporcov, Tatiana Natasha ^[5] ; Gama, Ricardo Ribeiro ^[6] ; Tajara, Eloiza Elena ^[7] ; Nunes, Fabio Daumas ^[1]

Affiliation

^[1] Univ Sao Paulo, Sch Dent, Dept Oral Pathol, Ave Prof Lineu Prestes 2227, BR-05508000 Sao Paulo - Brazil

^[2] Nove de Julho Univ UNINOVE, Postgrad Program Biophoton Appl Hlth Sci, BR-01504000 Sao Paulo - Brazil

^[3] Bauru Sch Dent, Dept Biol Sci, BR-17012901 Bauru - Brazil

^[4] Univ Sao Paulo, Sch Med, Dept Head & Neck Surg, BR-0317820 Sao Paulo - Brazil

^[5] Univ Sao Paulo, Sch Publ Hlth, BR-03178200 Sao Paulo - Brazil

^[6] Barretos Canc Hosp, Dept Head & Neck Surg, BR-01478440 Barretos - Brazil

^[7] Sch Med Sao Jose do Rio Preto, Dept Mol Biol, BR-15090000 Sao Jose Do Rio Preto - Brazil

• Document type: Journal article
• Source: International Journal of Oncology; v. 53, n. 6, p. 2458-2472, DEC 2018.
• Web of Science Citations: 1

Abstract

Oral squamous cell carcinoma (OSCC) is an extremely aggressive disease associated with a poor prognosis. Previous studies have established that cancer stem cells (CSCs) actively participate in OSCC development, progression and resistance to conventional treatments. Furthermore, CSCs frequently exhibit a deregulated expression of normal stem cell signalling pathways, thereby acquiring their distinctive abilities, of which self-renewal is an example. In this study, we examined the effects of GLI3 knockdown in OSCC, as well as the differentially expressed genes in CSC-like cells (CSCLCs) expressing high (CD44(high)) or low (CD44(low)) levels of CD44. The prognostic value of GLI3 in OSCC was also evaluated. The OSCC cell lines were sorted based on CD44 expression; gene expression was evaluated using a PCR array. Following this, we examined the effects of GLI3 knockdown on CD44 and ESA expression, colony and sphere formation capability, stem-related gene expression, proliferation and invasion. The overexpression of genes related to the Notch, transforming growth factor (TGF)beta, FGF, Hedgehog, Wnt and pluripotency maintenance pathways was observed in the CD44(high) cells. GLI3 knockdown was associated with a significant decrease in different CSCLC fractions, spheres and colonies in addition to the downregulation of the CD44, Octamer-binding transcription factor 4 (OCT4; also known as POU5F1) and BMI1 genes. This downregulation was accompanied by an increase in the expression of the Involucrin (IVL) and S100A9 genes. Cellular proliferation and invasion were inhibited following GLI3 knockdown. In OSCC samples, a high GLI3 expression was associated with tumour size but not with prognosis. On the whole, the findings of this study demonstrate for the first time, at least to the best of our knowledge, that GLI3 contributes to OSCC stemness and malignant behaviour. These findings suggest the potential for the development of novel therapies, either in isolation or in combination with other drugs, based on CSCs in OSCC. (AU)

• FAPESP's process: 12/00786-1 - Functional analysis of cell signaling involvement in cancer stem cells obtained from oral squamous cell carcinoma cell lines
• Grantee: Fabio Daumas Nunes
• Support Opportunities: Regular Research Grants
• FAPESP's process: 11/21395-8 - Functional analysis of cell signaling involvement in cancer stem cells obtained from oral squamous cell carcinoma cell lines
• Grantee: Maria Fernanda Setúbal Destro Rodrigues
• Support Opportunities: Scholarships in Brazil - Post-Doctoral