Aktivitas Anti Kanker Ekstrak Daun Afrika (Vernonia Amygdalina Delile) Secara In Vitro

Abstract

Pancreatic cancer is one type of cancer that continues to develop in the world. African leaf plants (Vernonia amygdalina Delile) which contain compounds such as sesquiterpenes lactone have the potential to be developed into anti-pancreatic cancer drugs. Proof of African leaves as an anticancer pancreas is carried out by cytotoxic test stages, cell cycle inhibition, apoptosis induction and inhibition of PI3K/Akt/mTOR gene expression. This test aims to determine the anticancer activity of african leaves extracts on pancreatic cancer cells (PANC-1). African leaves n-hexane extract (ALnHE), African leaves ethyl acetate extract (ALEAE) and African leaves ethanol extract (ALEE) were tested for cytotoxic activity against PANC-1 cells using the MTT assay method. The extract with the lowest IC50 value was tested on vero cells so that the selectivity index (IS) value was obtained. Cell cycle inhibition was carried out by the flow cytometry method, while the induced in cell apoptosis with the double staining method. The inhibition of PI3K/Akt/mTOR gene expression was tested by the Reverse Transcriptase Poly Chain Reaction (RT-PCR) method. The results showed that ALEAE had the lowest IC50 value of 29.77 ± 0.60 μg/mL and the selectivity index was 5.21. ALEAE with concentration 15 μg/mL leaded inhibits the cell cycle in the G2-M phase with a barrier percentage of 31.61% and 29.67% concentration of 6 μg/mL. ALEAE was also capable of induced apoptosis and inhibited PI3K/Akt/mTOR gene expression with a percentage of inhibition of 0.77 ± 0.00; 0.74 ± 0.01; and 0.71 ± 0.01 at a concentration of 15 μg/mL, whereas the concentration of 6 μg/mL were 0.85 ± 0.00; 0.77 ± 0.00; and 0.71 ± 0.01 respectively. Based on the explanation above it can be concluded that ALEAE has anticancer activity on pancreatic cancer cells PANC-1, anticancer mechanism through cell cycle inhibition, induces apoptosis and inhibition of PI3K/Akt/mTOR gene expression.

Kanker pankreas merupakan salah satu jenis penyakit kanker yang terus berkembang di dunia. Tumbuhan daun afrika (Vernonia amygdalina Delile) yang mengandung senyawa seperti seskuiterpen lakton dan glikosida berpotensi untuk dikembangkan menjadi obat antikanker pankreas. Pembuktian daun afrika sebagai antikanker pankreas dilakukan dengan tahapan uji sitotoksik, penghambatan siklus sel, pemacuan apoptosis dan penghambatan ekspresi gen PI3K/Akt/mTOR. Pengujian ini bertujuan untuk mengetahui aktivitas antikanker ekstrak daun afrika terhadap sel kanker pankreas (PANC-1). Ekstrak n-heksana daun afrika (EnHDA), ekstrak etil asetat daun afrika (EEADA) dan ekstrak etanol daun afrika (EEDA) hasil maserasi bertingkat aktivitas sitotoksiknya diuji terhadap sel PANC-1 dengan metode MTT assay. Ekstrak dengan nilai IC50 terendah diujikan terhadap sel vero sehingga diperoleh nilai indeks selektifitas (IS). Penghambatan siklus sel dilakukan dengan metode flow sitometri, sedangkan peningkatan apoptosis sel dengan metode double staining. Penghambatan ekspresi gen PI3K/Akt/mTOR diuji dengan metode Reverse Transcriptase Poly Chain Reaction (RT-PCR). Hasil penelitian menunjukan bahwa EEADA memiliki nilai IC50 terendah sebesar 29,77 ± 0,60 μg/mL dan indeks selektivitas sebesar 5,21. EEADA dengan konsentrasi 15 μg/mL menghambat siklus sel pada fase G2-M sebesar 31,61% dan 29,67% pada konsentrasi 6 μg/mL. EEADA juga memacu apoptosis serta menghambat ekspresi gen PI3K/Akt/mTOR dengan persentase penghambatan masing-masing sebesar 0,77 ± 0,00; 0,74 ± 0,01; dan 0,71 ± 0,01 pada konsentrasi 15 μg/mL, sedangkan pada konsentrasi 6 μg/mL masing-masing sebesar 0,85 ± 0,00; 0,77 ± 0,00; dan 0,71 ± 0,01. Berdasarkan pemaparan di atas dapat disimpulkan bahwa EEADA mempunyai aktivitas antikanker terhadap sel kanker pankreas PANC-1, mekanisme antikanker melalui penghambatan siklus sel, pemacuan apoptosis dan penghambatan ekspresi gen PI3K/Akt/mTOR.