RhoA/ROCKs signaling is increased by treatment with TKI-258 and leads to increased apoptosis in SCC-4 oral squamous cell carcinoma cell line.

Abstract

BACKGROUND:

This study evaluated the effect of treatment with TKI-258 on apoptosis, involving Rho GTPases and their effectors in SCC-4 cells of oral squamous cell carcinoma.

METHODS:

Markers of cell death and apoptosis were analyzed in control and TKI-258-treated SCC-4 cells by flow cytometry. The involvement of Rho GTPases and effectors in the induction of apoptosis by TKI-258 was evaluated by quantification of cleaved PARP. Also, gene expression analysis of those proteins was performed.

RESULTS:

The treatment with TKI-258 led to a significant increase in cell death (7-AAD) and apoptosis (annexin V and cleaved PARP). When Rho GTPases were stimulated with LPA and inhibited with toxin A Clostridium difficile, the percentage of apoptotic cells increased and decreased, respectively. A similar effect was found when the treatment was with TKI-258 combined with LPA and toxin A. Treatment with TKI-258 significantly increased RhoA gene expression, while RhoB, RhoC, Rac1, and Cdc42 decreased significantly. ROCKs inhibitors (Y-27632 and HA-1077) reduced apoptosis compared with control. TKI-258 combined with Y-27632 or HA-1077 led to an increase in apoptosis compared with inhibitors only. Treatment with TKI-258 led to an increase in ROCK1 and ROCK2 gene expression, and a decrease in PAK1 and PAK2 gene expression.

CONCLUSIONS:

TKI-258 stimulates apoptosis in SCC-4 cells of oral squamous cell carcinoma. Possibly, RhoA GTPase and their effectors ROCKs participate in the signaling pathway inhibited by TKI-258.

CLINICAL RELEVANCE:

Therapies with multi-target inhibitors, such as TKI-258, may be promising alternatives for the clinical treatment of oral squamous cell carcinoma.

• Authors: Anna Cecília Dias Maciel Carneiro, Fernanda Bernadelli De Vito, Helio Moraes-Souza, Virgínia Oliveira Crema
• Journal: Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology (J Oral Pathol Med) Vol. 50 Issue 4 Pg. 394-402 (Apr 2021) ISSN: 1600-0714 [Electronic] Denmark
• PMID: 33222274 (Publication Type: Journal Article)
• Copyright: © 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Chemical References

• 4-amino-5-fluoro-3-(5-(4-methylpiperazin-1-yl)-1H-benzimidazol-2-yl)quinolin-2(1H)-one
• Benzimidazoles
• Quinolones
• RHOA protein, human
• ROCK1 protein, human
• rho-Associated Kinases
• rhoA GTP-Binding Protein

Topics

• Apoptosis
• Benzimidazoles
• Carcinoma, Squamous Cell (drug therapy, genetics)
• Cell Line
• Head and Neck Neoplasms
• Humans
• Mouth Neoplasms (drug therapy, genetics)
• Quinolones
• Signal Transduction
• Squamous Cell Carcinoma of Head and Neck
• rho-Associated Kinases (genetics)
• rhoA GTP-Binding Protein (genetics)