Skripsi

Efek Pemberian Ekstrak Etanol Biji Delima (Punica granatum) terhadap Penghambatan Pertumbuhan dan Ekspresi Protein Inducible Nitric Oxide Synthase (iNOS) Sel Kanker Kolorektal HCT116 = The Effect of Ethanol Extract from Pomegranate (Punica granatum) Seed on Growth Inhibition and Expression of Inducible Nitric Oxide Synthase (iNOS) in Colorectal Cancer Cell Line HCT116.

Pendahuluan. Mortalitas akibat kanker kolorektal di Indonesia sebesar 9,5% dari seluruh kasus kanker. Pasien kanker kolorektal dengan ekspresi iNOS yang tinggi diketahui memiliki prognosis yang buruk. Sampai saat ini diketahui terdapat dua specific drug therapy dengan target EGFR dan VEGF yang dapat digunakan pada pasien kanker kolorektal. Akan tetapi, kedua obat tersebut masih terbatas penggunaannya serta menimbulkan efek samping. Delima (Punica granatum) diketahui sebagai tanaman herbal yang memiliki aktivitas antioksidan, antiinflamasi, dan antikanker. Beberapa penelitian telah menguji efek delima terhadap kanker. Akan tetapi, penelitian mengenai efek biji delima terhadap kanker masih minim. Metode. Aktivitas antikanker ekstrak etanol Punica granatum secara in vitro diuji menggunakan metode MTT assay pada cell line kanker kolorektal HCT116. Terdapat delapan serial konsentrasi yang diuji melalui MTT assay. Efek ekstrak etanol Punica granatum terhadap ekspresi protein iNOS pada cell line kanker kolorektal HCT116 dinilai melalui penghitungan nilai H-score dari pewarnaan imunositokimia. Terdapat empat kelompok perlakuan; kontrol negatif (0 ppm), dosis kecil (50 ppm), dosis sedang (100 ppm), kelompok dosis besar (200 ppm). Tiga konsentrasi dengan persentase inhibisi terbesar dari hasil MTT assay digunakan sebagai dosis esktrak pada imunositokimia. Hasil. Ekstrak etanol biji delima (Punica granatum) menunjukkan aktivitas antikanker melalui penghambatan pertumbuhan sel kanker kolorektal HCT116 dengan nilai IC50 sebesar 54,2763 µg/ml. Penurunan ekspresi protein iNOS dengan rerata nilai H-score sebesar 158,48 terjadi setelah diberikan ekstrak dengan dosis 200 ppm. Kesimpulan. Penelitian ini membuktikan bahwa biji delima (Punica granatum) menghambat pertumbuhan serta menurunkan ekspresi protein iNOS kanker kolorektal.
Kata kunci : IC50, imunositokimia, iNOS, kanker kolorektal, MTT assay, Punica granatum


Introduction. Mortality rate due to colorectal cancer in Indonesia was 9,5% from all cancer cases. Colorectal cancer patients that expressed iNOS in high amount known to have poor prognosis. There are two specific drug therapy targeted EGFR and VEGF for colorectal cancer treatment. Unfortunately, both of the drug can not be used for all patients and still have side effects. Pomegranate (Punica granatum) known to have antioxidant, anti-inflammatory, and anticancer activities. Various researches have been conducted to study the effect of pomegranate for cancer, but research that focus on pomegranate seed for cancer still minimal. Method. The in vitro anticancer activity of ethanol extract from Punica granatum seed was tested with MTT assay on HCT116 colorectal cancer cell line. The effect of ethanol extract from Punica granatum seed on iNOS expression studied by calculating the Hscore from immunocytochemistry staining. There were four groups; negative control (0 ppm), small dose (50 ppm), medium dose (100 ppm), and large dose (200 ppm). Three concentrations with highest percentage of inhibition from MTT assay were used as the dosage in this immunocytochemistry staining. Results. Ethanol extract from Punica granatum seed show anticancer activity by inhibiting HCT116 cell growth with IC50 of 54,2763 µg/ml. Ekspression of iNOS protein was decreased with mean of H-score 158,48 after administration of Punica granatum seed ethanol extract at a dose of 200 ppm. Conclusions. This study shows that Punica granatum seed can inhibit cancer growth and lowering iNOS expression in colorectal cancer cell line HCT116.
Keywords: colorectal cancer, IC50, immunocytochemistry, iNOS, MTT assay, Punica granatum