Abstract 3909: Defining Docetaxel’s antitumor effects on Calu3 lung cancer cells using high-throughput live-cell imaging

Abstract Introduction: Oncology focuses on understanding cancer's cellular mechanisms and developing effective treatments, with a key emphasis on evaluating drug toxicity to identify potential chemotherapies. Various assays, including cell viability, migration, and clonogenic assays, are used to assess the sensitivity and resistance of cancer cells to these drugs and define the drug’s profile. Docetaxel, a chemotherapeutic agent inhibiting microtubule depolymerization, involved in migration and proliferation, is studied using these assays to evaluate its impact on cell proliferation, survival, and migration. The results of toxicity studies can vary significantly depending on the type of assay used, and combining multiple assays enhances the accuracy of drug profiling. In this study, the Omni Pro 12 high-throughput live-cell imaging platform was used to simultaneously perform multiple assays on Calu3 lung cancer cells. Method: By conducting the assays simultaneously using the Omni Pro 12, variability was minimized, ensuring that differences in experimental conditions cannot affect the results. The aim was to evaluate the effects of Docetaxel on three key aspects of cancer progression: cell viability, migration, and colony-forming potential. These aspects were investigated using the cytotoxicity, scratch, and colony assay with Docetaxel concentrations between 0 - 1000 nM. Results: First, real-time cell confluence measurements indicated that higher Docetaxel concentrations (500 nM to 1000 nM) significantly suppressed cell proliferation. Secondly, a wound-healing assay revealed a dose-dependent inhibition of cell migration, with higher drug concentrations resulting in slower and an overall decrease in scratch closure. A significant 70% decrease in scratch closure was seen when comparing the control group to 1000 nM Docetaxel. Lastly, a clonogenic assay showed a dose-dependent decrease in colony formation at 324 hours post-treatment, the time point was chosen right before the control group colonies clustered, with higher concentrations (100 nM and 250 nM) leading to inhibition of colony growth with less than 10 colonies formed compared to approximately 110 in the control groups. Conclusion: These results demonstrate that Docetaxel effectively inhibits Calu3 cancer cell viability, proliferation, and migration in a dose-dependent manner. A comprehensive toxicity profile was obtained with the use of multiple 2D in vitro assays, which offers valuable insights into the potential therapeutic efficacy of Docetaxel in lung cancer treatment. This study underscores the importance of integrating multiple assays to better predict drug responses, helping to reduce the incidence of failures during in vivo and preclinical studies. Utilizing the Omni Pro 12 reduced the variance between assays as they are all performed simultaneously with similar experimental conditions. Citation Format: Skylar Dewees, Linda Boekestijn, Ben Streeter, Stacie Chvatal, Danielle Califano. Defining Docetaxel’s antitumor effects on Calu3 lung cancer cells using high-throughput live-cell imaging [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3909.