Cytotoxic and MMPs inhibitory activities of Sappan Wood (Caesalpiniasappan L.): various extracts on 4T1 breast cancer cell line
Abstract
Latar belakang: Kayu secang (Caesalpinia sappan L.) merupakan salah satu tanaman potensial dengan berbagai khasiat obat, termasuk antikanker. Gelatinase (MMP-2 dan MMP-9) merupakan matriks metaloproteinase (MMPs) dan memiliki peran penting dalam inisiasi, invasi, dan metastasis kanker. Penelitian ini bertujuan untuk menguji aktivitas sitotoksik dan penghambatan MMP dari ekstrak kayu secang pada sel kanker payudara 4T1.
Metode: Serbuk kayu secang dibagi menjadi 4 bagian, masing-masing diekstraksi dengan pelarut berbeda. Etanol 96%, etanol 70%, dan metanol digunakan untuk metode maserasi, sedangkan air untuk metode infusa. Uji MTT digunakan untuk menentukan efek sitotoksisitas, dan uji gelatin zymography untuk mendeteksi aktivitas MMP-2 dan MMP-9. Profil fitokimia ekstrak diamati dengan Kromatografi Lapis Tipis (KLT).
Hasil: Profil fitokimia ekstrak air menunjukkan profil KLT yang berbeda, sementara tiga ekstrak lainnya memiliki profil yang sama. Ekstrak etanol 96% kayu secang memberikan efek sitotoksik paling kuat terhadap 4T1 dengan nilai IC50 13,1 μg/mL, diikuti metanol (21,4 μg/mL), etanol 70% (22,5 μg/mL) dan air (25,5 μg/mL). Analisis gelatin zymograph dengan software ImageJ menunjukkan bahwa semua ekstrak kecuali air, menghambat aktivitas gelatinolitik MMP-9
Kesimpulan: Ekstrak kayu secang dengan berbagai pelarut polar memiliki aktivitas sitotoksik dan penghambatan MMP pada sel kanker payudara 4T1 dan berpotensi untuk dikembangkan sebagai anti kanker payudara.
Kata kunci: Caesalpinia sappan, MMP, sitotoksik, 4T1
Abstract
Background: Sappan wood (Caesalpinia sappan L.) is one of potential plant with wide variety of medicinal properties, including anticancer. Gelatinases (MMP-2 and MMP-9) are the member of matrix metalloproteinases (MMPs) and having a key role in cancer initiation, invasion, and metastasis. This study evaluated the cytotoxic and MMPs inhibitory activities of sappan wood in various extract on 4T1 breast cancer cell lines.
Methods: Sappan wood powder were divided into 4 parts, each part was extracted with different solvent. Ethanol 96%, ethanol 70%, and methanol were used for maceration methods, while water for infusion method. MTT assay used to identify cytotoxicity effect, and gelatin zymography assay to detect the activity of MMP-2 and MMP-9. Phytochemical profiling of the extract were observed by Thin Layer Chromatography (TLC).
Results: Phytocemical profiling of water extract showed different TLC profile, while three others had similar profile. The results of MTT assay showed that ethanolic 96% extract exhibited the strongest cytotoxic effect against 4T1 with the IC50 value 13,1 μg/mL, followed by methanolic (21,4 μg/mL), ethanolic 70% (22,5 μg/mL) and water (25,5 μg/mL). The analysis of gelatin zymograph bands using ImageJ software proved that all extracts except water, inhibited gelatinolytic activity of MMP-9.
Conclusion: The results of the study suggest that various extract of the sappan wood have been found to posses cytotoxic and MMPs inhibitory activities.
Keywords: Caesalpinia sappan, MMP’s, cytotoxic, 4T1
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