Induksi opoptosis galur sel osteosarkoma mg-63 oleh asam kafeat melalui caspase secara spesifik : Analisis sub-g1
A sam kafeat adalah senyawa fenolik sederhana yang banyak ditemukan pada tanaman hijau, buah, rempah, dan kopi. Asam kafeat memiliki berbagai efek biologis, salah satunya adalah potensinya sebagai senyawa anti kanker. Sebuah penelitian terdahulu telali membuktikan baliwa asam kafeat dapat menginduksi apoptosis pada sel osteosarkoma MG-63 serta menunjukkan adanya keterlibatan caspase 8, 9 dan 3 dalam mekanisme molekidamya. Nanmn, peran jalur pensinyalan caspase tersebut masih belum diketahui secara spesifik. Penelitian ini bertujuan imtuk membuktikan bahwa asam kafeat dapat menginduksi apoptosis pada galur sel osteosarkoma MG-63 secara spesifik melalui jalur pensinyalan caspase. Sel MG-63 dikultur dan dibagi menjadi 3 kelompok. Pada kelompok pertama, sel MG-63 diberikan pelarut sebanyak voliune inhibitor selama 2 jam, lalu diberikan pelarut sebanyak volume asam kafeat selama 48 jam. Pada kelompok kedua, sel MG-63 diberikan pelarut sebanyak volume inhibitor selama 2 jam, diikuti dengan pemberian asam kafeat dengan konsentrasi lOpg/mL selama 48 jam. Pada kelompok ketiga, kultur sel MG-63 ketiga diberikan Benzyloxycarbonyl-Val-Ala-Asp(0-methyl)-fluoromethylketone (Z-VAD-FMK) selama 2 jam, lalu diberikan asam kafeat selama 48 jam. Setelah perlakuan, sel diberikan solusi fluorokrom hipotonik yang mengandung propidium iodide, sodium sitrat, dan Triton X-100 selama 4 jam dan diukur menggunakan flow cytometer pada area Snb-Gl. Data yang diperoleh menunjukkan terdapat perbedaan signifikan pada persentase sel apoptosis antara kelompok pertama dengan kedua (£>=0,001), kelompok kedua dan ketiga (p=0,001), serta kelompok pertama dengan ketiga (£>=0,001). Hasil penelitian menunjukkan baliwa asam kafeat berpotensi sebagai anti osteosarkoma. dengan menginduksi apoptosis melalui jalur pensinyalan caspase secara spesifik pada sel kanker osteosarkoma.
C affeic acid is a simple phenolic compound, widely found in vegetables, fruits, spices and coffee. Anti-cancer is one of its highlighted biological effects. A previous study has shown that caffeic acid could induce apoptosis in MG-63 osteosarcoma cell line and indicates the involvement of caspase 8, 9 and 3. However, specific role of caspase is is still unclear. Therefore this study was conducted. MG-63 cells were cultured and divided into three groups. For the first group, MG-63 cells were treated with addition of solvent as much as inhibitor volume for 2 hours, followed by addition of solvent as much as caffeic acid volume for 48 hours. For the second group, MG-63 cells were treated with addition of solvent as much as the inhibitor for 2 hours, followed by addition of caffeic acid for 48 hours. For the third group, MG-63 cells were treated with addition of Benzyloxycarbonyl-Val-Ala-Asp(0-methyl)-fluoromethylketone (Z- VAD-FMK) for 2 hours, followed by addition of caffeic acid for 48 hours. Cells were then treated with hypotonic fluorocrom solution containing propidium iodide, sodium citrate, and Triton X-100 for 4 hours and quantified with flow cytometer at the sub-Gl area. Data obtained showed significant differences in the percentages of apoptotic cells between the first and the second groups (p=0.001), the second and the third groups (p=0.001), as well as the first and the third groups (p=0.00T). In conclusion, our results suggested that caffeic acid has potential as an anti-osteosarcoma by inducing apoptosis through specific caspase signaling pathway in osteosarma cancer cell.