Investigating the cytotoxic effects of AZD5363 on lung cancer cells

Abstract

Background: Lung cancer is reported to be the most commonly occurring cancer in the world, being the leading cause of cancer death among men, and the second most common cause of death among women after breast cancer (Sung, Ferlay et al. 2021). The PI3K/Akt/mTOR pathway has been strongly linked to the development and progression of NSCLC, which constitutes 85% of all lung cancers (Cheng, Shcherba et al. 2014, Papadimitrakopoulou 2012). This pathway is therefore a compelling target for anticancer therapy in NSCLC due to the extensive downstream signalling effects it has on the onset and progression of malignancy, as well as its potential impact on response and resistance to conventional therapies (Tan 2020). Among the PI3K/Akt/mTOR inhibitors is AZD5363 – a very potent and orally bioavailable pyrrolopyrimidine-derived pan-Akt kinase inhibitor which competes with ATP for Akt kinase association at the ATP binding site (Andrikopoulou, Chatzinikolaou et al. 2022). Besides the ongoing National Lung Matrix Trial (Middleton, Fletcher et al. 2020), few other investigations have, to date, tested the efficacy of this drug in different types of lung tumours. Aim: This study therefore aimed to investigate AZD5363 by sensitising two forms of aggressive NSCLC cells. Methodology: PrestoBlueTM cell viability assays were carried out to test the cytotoxicity of varying concentrations of AZD5363 on H520 (derived from squamous cell carcinoma) and A549 (originating from adenocarcinoma) cell lines. With the identified concentration that resulted in a 30% loss in cell viability, an ELISA assay was performed on A549 cells, to quantify how much of the protein eIF4E was being decreased. The same drug concentration was also utilised for wound healing assays, to investigate cell migratory behaviour of A549 cells in response to drug treatment. Results: The treatment of 10 – 100 μM AZD5363 on H520 cells failed to achieve a 30% loss in cell viability, and therefore no further investigations were conducted on this cell line. With respect to A549 cells, the same treatment decreased cell viability in a concentration-dependent fashion, such that 55 μM AZD5363 was selected as the 30% inhibitory concentration (IC30) for the subsequent investigations carried out on this cell line. In the ELISA assay, treatment of A549 cells with 55 μM AZD5363 achieved a reduction of 29% in mean eIF4E concentration. Following wound healing assays, percentage wound closure of A549 cells treated with 55 μM AZD5363 reached 33% after 72 hours of treatment, compared to 11% reached by cells treated with vehicle control solution. Conclusion: The reduction in mean eIF4E concentration achieved with 55 μM AZD5363 (IC30), as well as the migratory inhibition observed, indicate that this drug may be further investigated in combination with chemotherapy or other investigational treatment regimens, to potentially improve survival and quality of life.