Comparison of mtor and mlst8 moleculars with immunoflorence method in prostate cancer ctem cell culture with prostate cancer cell culture without stem cell and role of mtor pathway in prostate cancer

Background and Aim: Prostate cancer is the second most common cancer-related death in men. Investigation of the characteristics of cancer stem cells and the identification of the pathways involved in the development of cancer are important in the development of targeted treatment modalities. The mTOR pathway is actively involved in prostate cancer. The mTOR and mLST8 molecules are found in both the mTOR1 and mTOR2 complexes. The aim of this study is to compare the intensity of mTOR and mLST8 molecules in stem cell-free (Du-145 non-CSC) prostate cancer cell line culture with stem cell-containing prostate cancer cell culture (Du-145 CSC) by immunofluorescence. Materials And Methods: Prostate cancer cell line (Du-145) was cultured in an incubator at 37°C in 5% CO2 environment in cell culture flasks. Using flow cytometry analysis with fluorescence activated cell sorting (FACS), the cancer stem cells containing CD133+/ CD44+/low surface markers (sorting) in the Du-145 prostate cancer cell line and the remaining cells (non-sorting) were separated into 5 ml polystyrene tubes which includes 2 ml RPMI 1640 medium. Du-145 CSC and Du-145 non-CSCs were visually analyzed for both cell viability (DAPI method) and the intensity of fluorescence in mTOR and MLST8 proteins using the immunofluorescence staining method. The t-test was used as a statistical method and p<0.05 was considered statistically significant. Results: Changes in mTOR and MLST8 proteins in Du-145 CSC and Du-145 non-CSCs were examined by immunofluorescence staining. When compared to DU-145 non-CSC, there was an increase in fluorescence of mTOR and MLST8 proteins in DU-145 CSC. Intensity values in the immunofluorescence images of mTOR and mLST8 were calculated using the ImageJ program, an image analysis program. The density values of immunofluorescence images of mTOR protein in Du-145 CSC and Du-145 non-CSCs were compared. The density values of mTOR and MLST8 proteins in the prostate cancer stem cell were statistically significant. Conclusions: This significant increase in the mTOR and MLST8 proteins in the prostate cancer root cell may be an important part of the treatment and prognosis determination. It is possible to develop targeted therapies by conducting studies on mTOR pathway. Keywords: Prostate Cancer cell line (Du-145);Cancer Stem Cell; mTOR;MLST8