Investigation of the antiproliferative effects of docetaxel in combination with WNT signaling pathway inhibitor FH535 in non-small cell lung cancer

In our study, the effects of combined use of the WNT signaling pathway inhibitor FH535 and DTX used in the treatment of NSCLC, on adenocarcinomic human alveolar basal epithelium cell A549 and healthy bronchial epithelial cell BEAS-2B cell lines were evaluated by MTT and real-time cell analysis. While A549 reduces the activity of cancer cells, it is aimed to determine the combined dose showing the minimum damage in BEAS-2B healthy lung cell and to investigate the effects of this combined dose at the cellular level in vitro conditions with mitotic activity analysis, BrdU analysis and caspase 3-7 activity analysis methods in NSCLC. In this context, when giving minimum damage to the healthy lung cell; with the determination of the dose showing antiproliferative effect in NSCLC cell, it is aimed to establish new treatment protocols in addition to existing treatment protocols. According to the results we obtained; A combination dose of 0.005 μM DTX+0.5 μM FH535 showed anti-mitotic effects in the NSCLC cell line A549, causing mitotic catastrophism; it has been found to cause minimal damage to the healthy lung cell BEAS-2B and no more harm than the cytotoxic effect given in the use of DTX alone. This combination dose was found to have a high synergistic effect with a combination index value of 0.37505, showing a higher effect than the effect given by DTX and FH535 doses given separately. Therefore, the combined dose applied to the A549 cell line has been shown to be more effective than the use of both agents alone. When all the results are evaluated together, the proliferation rate in the A549 cell line was significantly reduced after the combined application. In real-time cell analysis, it was observed that the anti-mitotic activity of the cell increased and it died through mitotic catastrophe. Mitotic activity analysis results also supported the mitotic catastrophe result obtained from real-time cell analysis, and a decrease in mitotic activity was observed at 72 hours compared to the control group. It was investigated how the cell proliferation of the cell cycle in the synthesis phase was affected after the combined application with BrdU analysis. It was observed BrdU activity that decreased to 68% in 72 hours compared to the control group depending on time. As a result of, is an indication that the number of proliferated cells decreases with the reduction of mitotic division in parallel with the results of mitotic activity analysis. Caspase 3,7 activity was examined to show whether the cell died using the apoptotic cell death pathway after combined dose administration due to the decrease in cell viability and mitotic activity. There was not significant change in activity compared to the control group. This result, in parallel with the data obtained from real-time cell analysis, suggest that cancer cell treated with combined dose does not die using the apoptotic cell death pathway, and thus supports the idea that it goes to death using the mitotic catastrophe pathway within anti-mitotic effect. In our study, the effects of DTX and FH535 combination on NSCLC model A549 cell line and BEAS-2B healthy cell line were shown for the first time.