Immunofluorescence-Based Assay for High-Throughput Analysis of Multidrug Resistance Markers in Non-Small Cell Lung Carcinoma Patient-Derived Cells
2023
Аутори:
Dinić, JelenaPodolski-Renić, Ana
Dragoj, Miodrag
Jovanović Stojanov, Sofija
Stepanović, Ana
Lupšić, Ema
Pajović, Milica
Jovanović, Mirna
Petrović Rodić, Dušica
Marić, Dragana
Ercegovac, Maja
Pešić, Milica
Тип документа:
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт:
Lung cancer remains the leading cause of cancer death globally, with non-small cell lung cancer (NSCLC) accounting for the majority of cases. Multidrug resistance (MDR), often caused by ATP-binding cassette (ABC) transporters, represents a significant obstacle in the treatment of NSCLC. While genetic profiling has an important role in personalized therapy, functional assays that measure cellular responses to drugs are gaining in importance. We developed an automated microplate-based immunofluorescence assay for the evaluation of MDR markers ABCB1, ABCC1, and ABCG2 in cells obtained from NSCLC patients through high-content imaging and image analysis, as part of a functional diagnostic approach. This assay effectively discriminated cancer from non-cancer cells within mixed cultures, which is vital for accurate assessment of changes in MDR marker expression in different cell populations in response to anticancer drugs. Validation was performed using established drug-sensitive (NCI-H460) and drug-resistant (NCI-H460/R) NSCLC cell lines, demonstrating the assay’s capacity to distinguish and evaluate different MDR profiles. The obtained results revealed wide-ranging effects of various chemotherapeutic agents on MDR marker expression in different patient-derived NSCLC cultures, emphasizing the need for MDR diagnostics in NSCLC. In addition to being a valuable tool for assessing drug effects on MDR markers in different cell populations, the assay can complement genetic profiling to optimize treatment. Further assay adaptations may extend its application to other cancer types, improving treatment efficacy while minimizing the development of resistance.
Кључне речи:
lung cancer; NSCLC; multidrug resistance; MDR markers; ABCB1; ABCC1; ABCG2; immunofluorescence assay; primary cell culturesИзвор:
Diagnostics, 2023, 13, 24, 3617-Финансирање / пројекти:
- TargetedResponse - Functional diagnostics in non-small cell lung carcinoma: A new concept for the improvement of personalized therapy in Serbian patients (RS-ScienceFundRS-Ideje-7739737)