Dados do Trabalho

Título

Adenoid cystic carcinoma cellular models: establishment, characterization, and determination of chemoresistance

Introdução

Adenoid cystic carcinoma (AdCC) is the second most common malignant tumor of the salivary gland. The treatment of AdCC remains a challenging issue, and currently, there is no consensus on the use of chemotherapy for these tumors. The establishment of experimental models and the standardization of a protocol to use fresh tumor tissue samples to evaluate chemo resistance in AdCC tumors will allow a deeper understanding of AdCC oncogenesis and better tailoring of therapies.

Objetivo

Establishment and characterization of AdCC cellular models and development of an in vitro chemoresistance platform for these tumors.

Métodos

Tumor tissue samples from patients diagnosed with AdCC and submitted to surgery were collected and mechanically dissociated, being the fragments obtained used for primary cell culture. The cell monolayer growing from the explants was subcultured and the cells were characterized by immunocytochemistry, proliferation, migration, and invasion assays. The primary culture was also used to test tumor resistance to different drugs (cytotoxic and target therapies) used in the systemic treatment of AdCC. This study was approved by the Research Ethics Committee (protocol number 2624/18D).

Resultados

Two in vitro models (ACC1 and ACC3) were established from male patients with high-grade AdCC, originating respectively from the parotid gland and palate. The cultured tissue explants resulted in cell growth, and after 5-7 days, a cell monolayer was successfully established. Immunocytochemistry experiments revealed a strong expression of epithelial and myoepithelial biomarkers, such as cytokeratins, p63, SMA, and vimentin. The expression of Ki-67 and c-kit was also observed. The proliferation assay demonstrated a high cell proliferation rate up to 72 hours and decreased cell viability at 96h for ACC1, and a high proliferation rate up to 96 hours for ACC3. Both cells presented migration ability; however, ACC3 presented higher invasion ability compared to ACC1 cells. Both cells were tested with varying concentrations of cytotoxic and target therapies to evaluate tumor resistance. The results revealed distinct patterns of drug response.

Conclusões

Our study successfully established and characterized in vitro models of AdCC, providing a better understanding of the biological behavior of these tumors. Furthermore, these in vitro models hold promise as tools for the determination of a repertoire of drugs that might be applied for a personalized medicine.

Palavras-chave

Adenoid cystic carcinoma. Cell culture. Chemoresistance.

Financiador do resumo

Adenoid Cystic Carcinoma Research Foundation

Área

Estudo Clínico - Tumores de Cabeça e Pescoço