Publicação
Inhibition of the redox function of APE1 as a potential strategy to improve the efficacy of cisplatin in non-small cell lung cancer cells
| Resumo: | Lung cancer (LC) is the leading cause of cancer-related deaths worldwide for both men and women. Non-small cell lung cancer (NSCLC) accounts for the majority of all LC cases and has a low survival rate mainly due to metastasis progression. The standard chemotherapy medication for this type of cancer is the classical platinum-based therapy, being cisplatin commonly used. Even though cisplatin is associated with slightly better survival rates, resistance to therapy often occurs, being this a major limitation to its clinical use. Elevated expression levels of APE1 have been correlated with more aggressive phenotypes and poor prognosis of NSCLC. Besides being a key DNA repair enzyme, APE1 also acts as a redox signaling protein, modulating the activation of several transcription factors related to cancer progression, thus making it an emerging drug target in cancer therapy. For this reason, the quinone derivative E3330 has been tested as a direct inhibitor of APE1’s redox function in different types of tumors. In this context, the aim of the present study was to assess the effects of E3330 per se or in combination with cisplatin in NSCLC cells in terms of cytotoxicity, cell cycle distribution and cell migration/invasion. Firstly, the human H1975 cells were exposed to E3330 (5-50 µM) and/or cisplatin (1-50 µM) for 72 h and cell viability was assessed using the crystal violet (CV) and MTS assays. Cisplatin clearly decreased cell viability in a time- and concentration-dependent manner, with IC50 values of 9.6 µM for CV and 15.9 µM for MTS. The co-incubation of E3330 (30 µM) and cisplatin (5, 10 and 20 µM) significantly decreased cell viability compared to cisplatin alone, for all the concentrations tested and for both CV and MTS assays. In terms of cell cycle distribution, cisplatin led to an increase in sub-G1 and S-phases and decreased G0/G1-phase, whereas the co-treatment with E3330 did not alter the profile displayed by cisplatin-treated cells. The effect of E3330 in the migration and invasion of cisplatin-treated cells was also observed at non-cytotoxic concentrations of both compounds. In fact, E3330 significantly reduced both collective (wound-healing assay, ~20%) and chemotactic (transwell assay, ~12%) migration of H1975 cells exposed to cisplatin when compared with H1975 control cells. Furthermore, the combination of these two compounds also had an impact on cell invasion processes by reducing chemoinvasion of control cells (~17%). Overall, these results pointed out E3330 as a promising compound to boost cisplatin therapy that warrants further investigation in NSCLC. |
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| Autores principais: | Manguinhas, Rita Catarina Álvaro |
| Assunto: | Cisplatin E3330 NSCLC Migration/invasion Cytotoxicity Teses de mestrado - 2019 |
| Ano: | 2019 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Lung cancer (LC) is the leading cause of cancer-related deaths worldwide for both men and women. Non-small cell lung cancer (NSCLC) accounts for the majority of all LC cases and has a low survival rate mainly due to metastasis progression. The standard chemotherapy medication for this type of cancer is the classical platinum-based therapy, being cisplatin commonly used. Even though cisplatin is associated with slightly better survival rates, resistance to therapy often occurs, being this a major limitation to its clinical use. Elevated expression levels of APE1 have been correlated with more aggressive phenotypes and poor prognosis of NSCLC. Besides being a key DNA repair enzyme, APE1 also acts as a redox signaling protein, modulating the activation of several transcription factors related to cancer progression, thus making it an emerging drug target in cancer therapy. For this reason, the quinone derivative E3330 has been tested as a direct inhibitor of APE1’s redox function in different types of tumors. In this context, the aim of the present study was to assess the effects of E3330 per se or in combination with cisplatin in NSCLC cells in terms of cytotoxicity, cell cycle distribution and cell migration/invasion. Firstly, the human H1975 cells were exposed to E3330 (5-50 µM) and/or cisplatin (1-50 µM) for 72 h and cell viability was assessed using the crystal violet (CV) and MTS assays. Cisplatin clearly decreased cell viability in a time- and concentration-dependent manner, with IC50 values of 9.6 µM for CV and 15.9 µM for MTS. The co-incubation of E3330 (30 µM) and cisplatin (5, 10 and 20 µM) significantly decreased cell viability compared to cisplatin alone, for all the concentrations tested and for both CV and MTS assays. In terms of cell cycle distribution, cisplatin led to an increase in sub-G1 and S-phases and decreased G0/G1-phase, whereas the co-treatment with E3330 did not alter the profile displayed by cisplatin-treated cells. The effect of E3330 in the migration and invasion of cisplatin-treated cells was also observed at non-cytotoxic concentrations of both compounds. In fact, E3330 significantly reduced both collective (wound-healing assay, ~20%) and chemotactic (transwell assay, ~12%) migration of H1975 cells exposed to cisplatin when compared with H1975 control cells. Furthermore, the combination of these two compounds also had an impact on cell invasion processes by reducing chemoinvasion of control cells (~17%). Overall, these results pointed out E3330 as a promising compound to boost cisplatin therapy that warrants further investigation in NSCLC. |
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