Uji Aktivitas Sitotoksik N-Phenylpyrazoline A terhadap Cell Line SKOV-3 Model Kanker Ovarium: Studi In Vitro dan In Silico
Maulana Sabrang Amirullah, Prof. Dr. Mustofa, Apt., M.Kes., Dr. drh. Pamungkas Bagus Satriyo, Ph.D.
2025 | Skripsi | PENDIDIKAN DOKTER
Latar belakang: Kanker ovarium merupakan keganasan yang berasal dari indung telur, dengan sebagian besar kasus didominasi oleh epithelial ovarian cancer. Berdasarkan data GLOBOCAN 2022, terdapat lebih dari 320.000 kasus baru dan lebih dari 200.000 kematian akibat kanker ovarium secara global, menjadikannya kanker ginekologi yang umum. Faktor penyebabnya bersifat multifaktorial, termasuk genetik, lingkungan, dan gaya hidup, namun kehamilan dan penggunaan kontrasepsi hormonal diketahui dapat menurunkan risikonya. Kebanyakan kasus kanker ovarium baru didiagnosis serta diterapi ketika memasuki stadium lanjut yang memerlukan kombinasi obat kemoterapi. Akan tetapi, hal ini tidak menunjukkan respons yang signifikan dan berujung pada resistensi pengobatan. Salah satu mekanisme utama kanker ovarium adalah persinyalan ERBB2/HER2. Senyawa N-phenylpyrazoline, yang memiliki potensi sebagai antikanker pada kanker serviks, sedang diteliti untuk menguji aktivitas sitotoksiknya terhadap cell line SKOV-3 sebagai model kanker ovarium.
Tujuan: Mengkaji efek sitotoksik N-phenylpyrazoline A terhadap pertumbuhan cell line SKOV-3 secara in vitro dan menentukan protein target dari N-phenylpyrazoline A serta korelasi ekspresi protein target terhadap overall survival ratepasien kanker ovarium secara in silico.
Metode: Penelitian dilakukan secara in vitro dengan menggunakan tujuh tingkatan konsentrasi N-phenylpyrazoline A (1.25 µM, 2.5 µM, 5 µM, 10 µM, 20 µM, 40 µM, dan 80 µM) pada cell line SKOV-3. Analisis regresi nonlinier dilakukan untuk mendapatkan nilai IC50 dari N-phenylpyrazoline A dalam menghambat pertumbuhan sel. Secara in silico, situs Swiss Target Prediction digunakan untuk memprediksi protein target dari N-phenylpyrazoline A. Selain itu, Swiss Dock dan One Click Docking Mcule digunakan untuk melakukan uji molecular docking dan mengestimasi kekuatan pengikatan N-phenylpyrazoline A dengan protein target. Analisis Kaplan-Meier dilakukan pada data ketahanan hidup pasien kanker ovarium yang didapatkan dari UCSC Xena.
Hasil: N-phenylpyrazoline A memiliki aktivitas sitotoksik terhadap cell line SKOV-3 model kanker ovarium dengan nilai IC50 sebesar 58.29 M. Analisis protein target menunjukkan ERBB2, EGFR, PTK2, KDR, dan PRKCZ sebagai lima protein target dengan probabilitas terbesar. Uji molecular docking secara in silico menunjukkan bahwa N-phenylpyrazoline A memiliki afinitas yang lebih lemah dibandingkan dengan ligan standarnya yaitu lapatinib. Analisis public dataset menunjukkan bahwa hanya ekspresi ERBB2 yang berkorelasi secara signifikan terhadap overall survival rate yang lebih rendah dari pasien kanker ovarium (p < 0>
Kesimpulan: N-phenylpyrazoline A memiliki aktivitas sitotoksik yang sedang terhadap cell line SKOV-3 dengan target utama ERBB2 yang berkorelasi secara signifikan terhadap kelangsungan hidup pasien kanker ovarium yang lebih rendah.
Background: Ovarian cancer is a malignancy originating from the ovaries, with most cases dominated by epithelial ovarian cancer. Based on GLOBOCAN 2022 data, there are more than 320,000 new cases and more than 200,000 deaths from ovarian cancer globally, making it a common gynecological cancer. The causative factors are multifactorial, including genetics, environment, and lifestyle, but pregnancy and the use of hormonal contraceptives are known to reduce the risk. Most cases of ovarian cancer are only diagnosed and treated when they are in an advanced stage requiring a combination of chemotherapy drugs. However, this does not show a significant response and leads to treatment resistance. One of the main mechanisms of ovarian cancer is ERBB2/HER2 signaling. The compound N-phenylpyrazoline, which has potential as an anticancer in cervical cancer, is being studied to test its cytotoxic activity against the SKOV-3 cell line as an ovarian cancer model.
Objective: To study the cytotoxic effect of N-phenylpyrazoline A on the growth inhibition of SKOV-3 cell lines in vitro and to determine the target protein of N-phenylpyrazoline A and the correlation of target protein expression with the overall survival rate of ovarian cancer patients in silico.
Method: The study was conducted in vitro using seven levels of N-phenylpyrazoline A concentration (1.25 µM, 2.5 µM, 5 µM, 10 µM, 20 µM, 40 µM, and 80 µM) in the SKOV-3 cell line. Nonlinear regression analysis was performed to obtain the IC50 value of N-phenylpyrazoline A in inhibiting cell growth. In silico, the Swiss Target Prediction site was used to predict the target protein of N-phenylpyrazoline A. In addition, Swiss Dock and One Click Docking Mcule were used to perform molecular docking tests and estimate the binding strength of N-phenylpyrazoline A with the target protein. Kaplan-Meier analysis was performed on the survival dataset of ovarian cancer patients obtained from UCSC Xena.
Result: N-phenylpyrazoline A has cytotoxic activity against the SKOV-3 cell line, an ovarian cancer model with an IC50 value of 58.29 ?M. Target protein analysis showed ERBB2, EGFR, PTK2, KDR, and PRKCZ as the five target proteins with the highest probability. In silico molecular docking tests showed that N-phenylpyrazoline A has a weaker affinity compared to its standard ligand, lapatinib. Analysis of public datasets showed that only ERBB2 expression was significantly correlated with the overall survival rate of ovarian cancer patients (p < 0>
Conclusion: N-phenylpyrazoline A has moderate cytotoxic activity against the SKOV-3 cell line with the main target being ERBB2 which is significantly correlated with lower survival of ovarian cancer patients.
Kata Kunci : N-phenylpyrazoline, kanker ovarium, SKOV-3, ERBB2, protein
