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AT-101, (-)- gossypolün enantiomeri olup, potent antikanser ajandır. Tümör nekrozis faktör ilişkili apoptozisi uyaran ligant (Apo2L/TRAIL) apoptozisi, tercihen kanser hücrelerinde uyarır. Amaç, AT-101 ile Apo2L/TRAIL'in tekli ve kombine dozlarının sitotoksik ve apoptotik etkilerinin insan meme kanseri hücre hatlarında (MCF-7 ve MDA-MB-231) araştırılması ve hangi apoptotik moleküllerin etkilendiğinin ortaya konmasıydı. Hücreler, 72 saat süresince, AT-101 (0.5-10?M) ve TRAIL (10-200ng/mL) ile inkübe edildi. XTT tabanlı canlılık testi kullanılarak sitotoksisite yüzdeleri hesaplandı. Apoptozis, histon-DNA fragmentasyonu ölçülerek kanıtlandı.MCF-7 ve MDA-MB-231 hücrelerinde IC50 değerleri sırasıyla, 4.64 ?M, 3.99 ?M (AT-101) ve 133 ng/mL, 93.2 ng/mL (TRAIL) idi. Sitotoksisite değerleri MCF-7 ve MDA-MB-231 hücrelerinde sırasıyla % 32.3, % 36.6 AT-101(2.5 µM) ve % 36.8, % 35.7 TRAIL (60 ng/mL) `di.60ng/mL TRAIL+ 2.5 ?M AT-101 kombinasyonunda MCF-7 hücrelerinde sinerjistik sitotoksisite % 78.2 (KI: 0.298) iken, MDA-MB-231 hücrelerinde % 83.2 (KI: 0.256)'di. 2.5 ?M AT-101 ile 80ng/mLTRAIL'in ardışık muamelesinde, sitotoksisite değerleri MCF-7 ve MDA-MB-231 hücrelerinde sırasıyla % 80.8 ve % 86.8'di.AT-101 ve TRAIL'in kombinasyonu, MDA-MB-231 hücrelerinde, Bax, Trail-R1, Kaspaz-3, Sitokrom C proteinlerinde artışa neden olurken, Pon2, Procaspase-3, Bcl-x, Bcl-2 ve Xiap proteinlerinde azalışa neden oldu. MCF-7 hücrelerinde, Bad, Trail-R1, Pon2, Fadd, Smac/Diablo, Sitokrom C proteinlerinde anlamlı artış, Ciap proteininde anlamlı azalış oldu.AT-101 ve TRAIL kombinasyonunun, MCF-7 ve MDA-MB-231 hücre kültürlerindeki sinerjistik apoptotik etkileri, ilk defa bu çalışmayla belirlendi. AT-101 is an (-)- enantiomer of gossypol with potent anticancer agent. Tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) induces apoptosis preferentially in cancer cells. The objective was to investigate the cytotoxic and apoptotic effects of single and combined doses of AT-101 and Apo2L/TRAIL on human breast cancer cell lines (MCF-7 and MDA-MB-231) and to find out what apoptotic molecules have been affected. The cells were incubated with AT-101 (0.5-10?M) and TRAIL (10-200ng/mL) for 72 hours. XTT-based viability assay was used to calculate the percentage cytotoxicity. Apoptosis was confirmed by measuring the histon-DNA fragmentation degree.The IC50 values of MCF-7 and MDA-MB-231 cells were 4.64 ?M, 3.99 ?M (AT-101) and 133 ng/mL, 93.2 ng/mL (TRAIL) respectively. The cytotoxicity for MCF-7 and MDA-MB-231 cells was 32.3, 36.6 % in AT-101(2.5 µM) and 36.8, 35.7 % in TRAIL (60 ng/mL) respectively.The synergistic cytotoxicity was 78.2 % (CI: 0.298) at MCF-7 cells while it was 83.2 % (CI: 0.256) at MDA-MB-231 cells at combination (60ng/mL TRAIL+2.5 ?M AT-101). In sequential treatment of 2.5 ?M AT-101 and 80 ng/mLTRAIL, cytotoxicity were 80.8 and 86.8 % for MCF-7 and MDA-MB-231 cell cultures respectively.The combined use of AT-101 and TRAIL resulted in an increase at Bax, Trail-R1, Caspase-3, Cytochrome C, proteins and a decrease in Pon2, Procaspase-3 , Bcl-x, Bcl-2 and Xiap proteins in MDA-MB-231 cell cultures. There was significant increase in Bad, Trail-R1, Pon2, Fadd, Smac/Diablo, Cytochrome C and a decrease in Ciap protein in MCF-7 cell cultures.The study is the first primary data on the synergistic apoptotic effects of AT-101 and TRAIL combination in MCF-7 and MDA-MB-231 cell lines. 75 |
