Penghambatan Perkembangan Kanker Hati HepG2 oleh Ekstrak Daun Afrika (Vernonia amygdalina Delile): Kajian Molekuler Secara In Vitro

Abstract

Kanker hati adalah tumor ganas yang menyebabkan kerusakan bentuk dan fungsi organ hati. Kanker hati umumnya merupakan perkembangan dari hepatitis kronis atau sirosis. Daun afrika mengandung saponin, flavonoid, tanin glikosida, dan steroid/triterpenoid.. Berdasarkan penelitian terdahulu senyawa ini dilaporkan memiliki efek sitotoksik terhadap beberapa jenis sel kanker. Penelitian ini bertujuan untuk mengetahui aktivitas antikanker ekstrak dan senyawa aktif daun afrika dalam penghambatan sisklus sel dan pemacuan apoptosis terhadap sel kanker pankreas (HepG2). Simplisia daun afrika diekstraksi dengan cara maserasi bertingkat dengan menggunakan pelarut n-heksan, etil asetat, dan etanol. Pengujian sitotoksik ekstrak n-heksan daun afrika, ekstrak etil asetat daun afrika, ekstrak etanol daun afrika, dan sorafenib sebagai kontrol positif menggunakan 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 19.91 ± 0.24 μg/mL dan indeks selektivitas sebesar 7.79. EEADA dengan konsentrasi 15 μg/mL menghambat siklus sel pada fase G2-M sebesar 11.33%. EEADA juga memacu apoptosis serta menghambat ekspresi gen PI3K/Akt/mTOR dengan persentase penghambatan masing-masing sebesar 0.55 ± 0.00; 0.25 ± 0.01; dan 0.54 ± 0.01 pada konsentrasi 15 μg/mL. Berdasarkan hasil di atas dapat disimpulkan bahwa EEADA mempunyai aktivitas antikanker terhadap sel kanker hati HepG2, mekanisme antikanker melalui penghambatan siklus sel, pemacuan apoptosis dan penghambatan ekspresi gen PI3K/Akt/mTOR.

Liver cancer is a malignant tumor that causes damage to the form and function of liver. Liver cancer is generally aprogression from chronic hepatitis or cirrhosis. The African leaves have been known to contain saponins, flavonoids, tannin glycosides, and steroids/triterpenoids. According to previous studies, this compound was reported to have a cytotoxic effect on several types of cancer cells. Therefore, this study aimed to determine the anticancer activity of extracts and active compounds of African leaves in inhibiting cell cycle and inducing apoptosis on liver cancer cells (HepG2). The African leaf simplicia was extracted by multilevel maceration method using n-hexane, ethyl acetate, and ethanol solvent). The cytotoxic tests of n-hexane extract of African leaves, ethyl acetate extract of African leaves, the ethanol extract of African leaves, and sorafenib as a positive control were carried out 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 19,91 ± 0,24 μg/mL and the selectivity index was 7.79. ALEAE with concentration 15 μg/mL leaded inhibits the cell cycle in the G2-M phase with a barrier percentage of 11.33%. ALEAE was also capable of induced apoptosis and inhibited PI3K/Akt/mTOR gene expression with a percentage of inhibition of 0.55 ± 0,00; 0.25 ± 0.01; dan 0.54 ± 0.01 at a concentration of 15 μg/mL. Based on the explanation above it can be concluded that ALEAE has anticancer activity on liver cancer cells HepG2, anticancer mechanism through cell cycle inhibition, induces apoptosis and inhibition of PI3K/Akt/mTOR gene expression.