DINI NURUL UTAMI, - (2025) POTENSI SITOTOKSIK ISOLAT ANDROGRAFOLID (Andrographis paniculata (Burm.F)) TERHADAP SEL KANKER PROSTAT DU-145 DAN PARU A-549 MELALUI JALUR EKSTRINSIK DENGAN METODE IN CELL WESTERN. Skripsi thesis, Sekolah Tinggi Farmasi Indonesia.
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Abstract
Kanker paru (A-549) dan prostat (DU-145) memiliki angka insiden dan mortalitas tinggi, sementara kemoterapi sering menimbulkan efek samping berat dan resistensi obat. Andrographis paniculata (Burm.F) mengandung andrografolid yang dilaporkan bersifat antikanker melalui induksi apoptosis. Penelitian ini mengevaluasi potensi isolat andrografolid dari Sekolah Tinggi Farmasi Indonesia dalam menginduksi apoptosis pada A-549 dan DU-145 menggunakan metode InCell Western (ICW). Nilai IC₅₀ andrografolid adalah 47,28 μg/mL (A-549) dan 49,35 μg/mL (DU-145) dengan variasi dosis tiap kelompok menggunakan ½×IC₅₀, 1×IC₅₀, 2×IC₅₀, dan 3×IC₅₀, serta doxorubicin (A-549) dan cisplatin (DU-145) sebagai kontrol positif. Tahapan ICW meliputi fiksasi, permeabilisasi, blocking, pemberian antibodi primer (caspase-9), dan deteksi antibodi sekunder berlabel inframerah, diikuti pembacaan sinyal menggunakan scanner Li-Cor. Hasil menunjukkan perubahan morfologi apoptosis secara dosis-dependent. Ekspresi caspase-9 meningkat pada kedua lini sel, dengan puncak aktivasi pada dosis tinggi. Sel kanker paru A-549 menunjukkan respons sejak dosis 1×IC₅₀ yaitu sebesar 47,28 μg/mL, sedangkan DU-145 memerlukan dosis ≥2×IC₅₀ yaitu sebesar 98,7 μg/mL untuk mendekati efektivitas kontrol positif. Aktivasi caspase-9 mengindikasikan keterlibatan jalur intrinsik, dengan kemungkinan kontribusi jalur ekstrinsik melalui mekanisme caspase-8–tBid–mitokondria. Kesimpulannya, isolat andrografolid berpotensi sebagai agen antikanker alami, terutama pada kanker paru, melalui induksi apoptosis jalur intrinsik dengan indikasi keterlibatan jalur ekstrinsik. ----- Lung cancer (A-549) and prostate cancer (DU-145) had high incidence and mortality rates, while chemotherapy often caused severe side effects and drug resistance. Andrographis paniculata (Burm. F) contained andrographolide, which had been reported to possess anticancer properties through apoptosis induction. This study evaluated the potential of andrographolide isolate from Sekolah Tinggi Farmasi Indonesia to induce apoptosis in A-549 and DU-145 cells using the InCell Western (ICW) method. The IC₅₀ values of andrographolide were 47.28 μg/mL for A-549 and 49.35 μg/mL for DU-145. Treatments were administered at ½×, 1×, 2×, and 3×IC₅₀, with doxorubicin (A-549) and cisplatin (DU-145) as positive controls. The ICW procedure included fixation, permeabilization, blocking, incubation with primary antibody (caspase-9), and detection with infrared-labeled secondary antibody, followed by signal reading using a Li-Cor scanner. The results showed dose-dependent apoptotic morphological changes. Caspase-9 expression increased in both cell lines, with peak activation at higher doses. A-549 cells responded significantly at 1×IC₅₀ (47.28 μg/mL), while DU-145 cells required ≥2×IC₅₀ (98.7 μg/mL) to approach the effectiveness of the positive control. Caspase-9 activation indicated the involvement of the intrinsic pathway, with possible contribution from the extrinsic pathway via the caspase-8–tBid– mitochondria mechanism. In conclusion, andrographolide isolate had potential as a natural anticancer agent, particularly against lung cancer, through intrinsic pathway apoptosis induction with indications of extrinsic pathway involvement.
| Item Type: | Thesis (Skripsi) |
|---|---|
| Uncontrolled Keywords: | A-549, Andrografolid, DU-145, In-Cell Western. ------ A-549, Andrographolide, DU-14, In-Cell Western. |
| Subjects: | Q Science > Q Science (General) R Medicine > RM Therapeutics. Pharmacology |
| Divisions: | Program Studi S1 Farmasi |
| Depositing User: | pustakawan - - |
| Date Deposited: | 30 Aug 2025 02:40 |
| Last Modified: | 30 Aug 2025 02:40 |
| URI: | http://repository.stfi.ac.id/id/eprint/2729 |
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