Analisis In Silico dan In Vitro Antikanker Senyawa Kalkon 3 terhadap Sel Kanker Ovarium SKOV-3 Kokultur Peripheral Blood Mononuclear Cells

Chiesa Salsabila, drh. Retno Murwanti, M.P., Ph.D.; apt. Muhammad Novrizal Abdi Sahid, M.Eng., Ph.D.

2025 | Tesis | S2 Ilmu Farmasi

Di Indonesia, kanker ovarium memiliki prevalensi tinggi sebesar 64?ri total kasus pada tahun 2020. Upaya terus dilakukan untuk mengembangkan agen terapeutik yang menargetkan sel kanker dan bersinergi dengan sel imun dalam Tumor Microenvironment (TME). Salah satu antikanker potensial, senyawa kalkon 3 (C15H12O), telah terbukti menghambat pertumbuhan kanker payudara, serviks, dan kolon. Pada penelitian ini, sel kanker ovarium SKOV-3 dikokultur dengan Peripheral Blood Mononuclear Cell (PBMC) untuk memodelkan sel-sel imun dalam TME, sehingga perlakuan senyawa kalkon 3 dapat dilihat dari sistem monokultur SKOV-3 dan kokultur SKOV-3/PBMC. Tujuan dari penelitian ini adalah menganalisis potensi senyawa kalkon 3 sebagai antikanker dalam sistem kokultur.

Metode in silico dilakukan melalui data mining dan identifikasi protein target senyawa kalkon 3, Protein-Protein Interaction (PPI), analisis profil Overall Survival (OS) pasien kanker ovarium dan level ekspresi gen pada sel SKOV-3, serta analisis molecular docking senyawa kalkon 3 dengan protein target. Validasi secara in vitro dilakukan melalui uji sitotoksisitas dengan MTT Assay dan WST-1 Assay, deteksi apoptosis menggunakan flowcytometer, dan analisis ekspresi gen target menggunakan qRT-PCR.

Hasil data mining menunjukkan bahwa terdapat 109 gen target senyawa kalkon 3 pada kanker ovarium SKOV-3, termasuk Epidermal Growth Factor Receptor (EGFR), Peroxisome Proliferator-Activated Receptor Gamma (PPARG), Histone Deacetylase 1 (HDAC1), dan JUN yang ekspresi tingginya menurunkan OS pasien kanker ovarium. Hasil molecular docking menunjukkan bahwa kalkon 3 menargetkan EGFR dan PPAR-? dengan skor docking -6,5709 dan -7,1097. Hasil analisis sitotoksisitas menunjukkan bahwa kalkon 3 memiliki nilai IC50 sebesar 86 ?M pada sistem kokultur. Hasil uji apoptosis menunjukkan bahwa pemberian senyawa kalkon 3 sebanyak 86 ?M pasca kokultur meningkatkan apoptosis sebesar 54,03%. Hasil analisis ekspresi gen menunjukkan bahwa pemberian senyawa kalkon 3 pasca kokultur meningkatkan ekspresi EGFR dan menurunkan ekspresi PPARG bergantung pada dosis. Berdasarkan hasil penelitian, dapat disimpulkan bahwa senyawa kalkon 3 berpotensi sebagai antikanker ovarium dengan mentarget EGFR dan PPAR-?.

In Indonesia, ovarian cancer has a high prevalence of 64% of total cases in 2020. Innovation continues to be carried out to develop therapeutic agents that target cancer cells and synergize with immune cells in the Tumor Microenvironment (TME). One of the potential anticancer agents, chalcone 3 compound (C15H12O), has been proven to inhibit the growth of breast, cervical, and colon cancer. In this study, SKOV-3 ovarian cancer cells were co-cultured with Peripheral Blood Mononuclear Cells (PBMC) to model immune cells in the TME, allowing the effects of chalcone 3 to be observed from both the SKOV-3 monoculture and the SKOV-3/PBMC co-culture systems. The aim of this research is to analyze the potential of chalcone 3 as an anticancer agent in the co-culture system.

The in silico method was conducted through data mining and identification of target proteins of chalcone 3, Protein-Protein Interaction (PPI), analysis of Overall Survival (OS) profiles of ovarian cancer patients and gene expression levels in SKOV-3 cells, as well as molecular docking analysis of chalcone 3 with target proteins. In vitro validation was conducted through cytotoxicity tests using MTT Assay and WST-1 Assay, apoptosis detection using flowcytometer, and target gene expression analysis using qRT-PCR.

Data mining results showed that there are 109 target genes of chalcone 3 in SKOV-3 ovarian cancer, including Epidermal Growth Factor Receptor (EGFR), Peroxisome Proliferator-Activated Receptor Gamma (PPARG), Histone Deacetylase 1 (HDAC1), and JUN, whose high expression levels decrease the OS of ovarian cancer patients. Molecular docking results show that chalcone 3 targets EGFR and PPAR-? with docking scores of -6.5709 and -7.1097. The results of the cytotoxicity analysis show that chalcone 3 has an IC50 value of 86 ?M in the co-culture system. The apoptosis test results indicate that the administration of chalcone 3 at 86 ?M post-co-culture increases apoptosis by 54.03%. The gene expression analysis results show that the administration of chalcone 3 post-co-culture increases EGFR expression and decreases PPARG expression in a dose-dependent manner. Based on the research results, it can be concluded that chalcone 3 has the potential to be an ovarian anticancer agent by targeting EGFR and PPAR-?.

Kata Kunci : Antikanker, kalkon 3, PBMC, SKOV-3, TME