The effects of sulindac on cell viability and cell cycle in pharynx and larynx cancer cells

Aim: The purpose of this study is to research the effects of sulindac treatment on cell viability, cell cycle and angiogenesis in the pharynx and larynx carcinoma cell lines. Material and method: In a cell culture media a pharynx carcinoma cell line (FaDu) and larynx carcioma cell line (Detroit 562) has been incubated and reproducted in an incubator that has the settings of 37 oC and 5% CO2, and then has been passaged. The dose of sulindac has been established as 200 µM by using IC50 trials. To count and examine the viability of the cells, 1x105 cells has been transferred to each tube. By adding 450 μL MUSE Count Vability Reagent onto the 50 μL of cell suspension, we prepared a diluated sample to be stained, and then the calculations has been made by using MUSE device. To evaluate the cell cycle, 1x105 cells has been transferred to each tube. 70% ethanol has been added to the tubes and they have been incubated at-20 oC for 3 hours. After the centrifugation of the tubes, 200 μL MUSE Cell Cycle Reagent has been added, and then they have been incubated at room temperature in the dark for 30 minutes before the evaluation in the MUSE device. The cell cultures has been made with the 24-well plate and drugs have been addet to wells afterwards. The fixation with 4% paraformaldehyde at room temperature for 20 minutes has been performed and washing and incubation procedures is followed the fixation. We added VEGFR antibodies and enterprated the solution for occurence of any coloring with 3,3 diaminibenzidine. We analyzed the ADAMTS1 antibodies with Western Blot technique. Results: We found that the cell viability of both pharynx carcinoma cells (FaDu) and larynx carcioma cells (Detroit 562) is decreased after the sulindac treatment. In larynx carcinoma cells, the cell cycle was paused at the G0/G1 phase and in pharynx carcinoma cells, it paused at the G2/M phase after the sulindac treatment. This findings showed that the sulindac treatment causes the cell cycle in the larynx and pharynx carcinoma cells to pause at the phases that are before cell division and slows the cells down in the way of transition to mitosis. We found that sulindac increase the VEGF in the larynx and pharynx carcinoma cells in the immunohistochemistry analyses and decrease the levels of ADAMTS1 in the Western Blot analyses. This suggests that sulindac increases the angiogenesis. We have showed that beneficial outcomes for the treatment of larynx and pharynx cancer can be obtained by using sulindac. The future studies that examines sulindac and/or other non-steroidal anti-inflamatory drugs are needed to support our findings. Keywords: Sulindac, cell cycle, cell viability, angiogenesis, larynx cancer, pharynx cancer