Kajian Molekuler Penghambatan Pertumbuhan Sel Kanker WiDr dengan Pemberian Ekstrak n-heksan Beberapa Spesies Daun Mangrove

Abstract

Colon cancer is cancer that attacks the colon and rectum who is one of the deadly diseases worldwide and Indonesia. Avicennia alba, Avicennia lanata and Avicennia marina leaves contain polyisoprenoid compounds that have the potential to be developed into colonic anticancer drugs. This test aims to evaluate the anticancer activity of mangrove leaves extracts against colon cancer cells (WiDr). This study used 3 types of mangrove leaves namely Avicennia alba, Avicennia lanata and Avicennia marina. Extraction was carried out by maceration method by using n-hexane solvent to obtain Avicennia alba mangrove leaf nhexane extract (EnHDAA), Avicennia lanata mangrove leaf n-hexane extract (EnHDAL) and Avicennia marina mangrove leaf n-hexane extract (EnHDAM), tested inhibitory activity of the cancer cell cycle were carried out using the flowcytometry method, while the induced in cell apoptotic with double staining method, The Inhibition of PI3K/Akt1/mTOR/EGFR/P53 gene expression was performed by the Reverse Transcriptase Poly Chain Reaction (RT-PCR) method. Theresult showed that EnHDAA with a concentration of 34 μg/mL inhibited the cell cycle in the S and G2-M phases by 10.51% and 22.05%. EnHDAL with a concentration of 62 μg/mL inhibits the cell cycle in the S and G2-M phases of 10.60% and 23.84%, and EnHDAM with a concentration of 35 μg/mL inhibits the cell cycle in the G0-G1 phase by 90.49%. EnHDAA with a concentration of 34 μg/mL accelerates apoptotic by 9.86%, EnHDAL with a concentration of 62 μg/mL promotes apoptotic by 58.91% and EnHDAM with a concentration of 35 μg/mL stimulates apoptotic by 21.88%. The three extracts were able to down regulated gene expression PI3K/Akt1/mTOR/EGFR and also up regulated P53 expression. Based on the explanation above data, it can be concluded that EnHDAA, EnHDAL, and EnHDAM have anticancer activity against WiDr cells, the third anticancer mechanism through inhibition of cell cycle, induces apoptotic and inhibition of PI3K/Akt1/mTOR/EGFR/P53 in WiDr cells.

Kanker kolon adalah kanker yang menyerang usus besar dan rektum yang merupakan salah satu penyakit mematikan di dunia dan Indonesia. Tumbuhan daun mangrove Avicennia alba, Avicennia lanata dan Avicennia marina yang mengandung senyawa polyisoprenoid yang berpotensi untuk dikembangkan menjadi obat antikanker kolon.. Penelitian ini bertujuan untuk mengevaluasi aktivitas antikanker ekstrak daun mangrove terhadap sel kanker kolon (WiDr). Penelitian ini menggunakan 3 jenis daun mangrove yaitu Avicennia alba, Avicennia lanata dan Avicennia marina. Ekstraksi dilakukan dengan metode maserasi menggunakan pelarut n-heksan sehingga diperoleh ekstrak n-heksan daun mangrove Avicennia alba (EnHDAA), Ekstrak n-heksan daun mangrove Avicennia lanata (EnHDAL) dan Ekstrak n-heksan daun mangrove Avicennia marina (EnHDAM). Aktivitas penghambatan siklus sel WiDr dilakukan dengan metode flow sitometri, peningkatan apoptosis sel diamati dengan metode double staining. Penghambatan ekspresi gen PI3K/Akt1/mTOR/EGFR/P53 dilakukan dengan metode Reverse Transcriptase Poly Chain Reaction (RT-PCR). Hasil penelitian menunjukkan bahwa EnHDAA dengan konsentrasi 34 μg/mL menghambat siklus sel WiDr pada fase S dan G2-M sebesar 10,51 % dan 22,05%. EnHDAL dengan konsentrasi 62 μg/mL menghambat siklus sel pada fase S dan G2-M sebesar 10,60 % dan 23,84%, sedangkan EnHDAM dengan konsentrasi 35 μg/mL menghambat siklus sel WiDr pada fase G0-G1 sebesar 90,49%. EnHDAA dengan konsentrasi 34 μg/mL memacu apoptosis sebesar 9,86%, EnHDAL dengan konsentrasi 62 μg/mL memacu apoptosis sebesar 58,91% dan EnHDAM dengan konsentrasi 35 μg/mL memacu apoptosis sebesar 21,88%. Ketiga ekstrak mampu menurunkan ekspresi gen PI3K/Akt1/mTOR/EGFR dan juga meningkatkan ekspresi P53. Berdasarkan data-data di atas dapat disimpulkan bahwa EnHDAA, EnHDAL, EnHDAM mempunyai aktivitas antikanker terhadap sel WiDr. mekanisme antikanker ketiga ekstrak adalah melalui penghambatan siklus sel, pemacuan apoptosis dan penghambatan ekspresi gen PI3K/Akt1/mTOR/EGFR/P53 pada sel WiDr.