Dados do Trabalho

Título

In vitro and in vivo models of germ cell tumors: seeking to overcome cisplatin resistance

Introdução

Most patients with germ cell tumors (GCTs) respond well to standard cisplatin therapy; however, 15-20% of patients experience relapse. Researchers commonly use in vitro and in vivo models, including the xenograft model, to overcome cisplatin resistance. Recently, our research group established a cisplatin-resistant cell line of GCTs, NTERA-2R, which has shown promise in identifying new therapeutic targets. However, further in vivo studies using human models are necessary for personalized medicine.

Objetivo

This study aims to establish a xenograft model derived from GCT cell lines and evaluate whether it conserves the same expression pattern found in cell lines.

Métodos

This project received approval from CEUA (registered under number 010/2020). For in vivo experiments, 1x106 parental NTERA-2 (NTERA-2P) or resistant NTERA-2 (NTERA-2R) cells were subcutaneously injected into athymic mice (Mus musculus). The mice were observed and weighed weekly. Upon reaching tumor volumes close to 2000 mm3, the animals were euthanized, and the tumors were collected. Total RNA was extracted, quantified using Nanodrop, and reverse transcribed into complementary DNA (cDNA). Next, real-time PCR was performed using the TaqMan® assay (Applied Biosystems) to evaluate the expression of the genes MGMT, BRCA1, CDH1 (E-cadherin), VIMENTIN, and CDH2 (N-cadherin), with the GAPDH gene used as an endogenous control.

Resultados

The results revealed that the resistant cell line exhibited higher expression levels of VIMENTIN and CDH2 compared to the parental cells, indicating an alteration that favors the mesenchymal phenotype concerning the epithelial-mesenchymal transition (EMT), a biological process associated with cancer progression, resistance, and metastasis. Additionally, there was a trend towards increased BRCA1 expression and decreased CDH1. These findings are consistent with the in vitro data, further supporting our observations.

Conclusões

Therefore, the xenograft model using cisplatin-resistant NTERA-2 cells partially preserved the gene expression pattern observed in vitro. The successful establishment of this in vivo model for GCTs represents a significant advancement and paves the way for future studies on chemotherapeutic agents, as well as contributes to the progress of personalized medicine.

Palavras-chave

1. Germ cell tumor
2. Drug resistance
3. Epithelial-mesenchymal transition

Financiador do resumo

Teaching and Research Institute of the Barretos Cancer Hospital. FAPESP Process n. 2019/07502-8.