Studi Pengaruh Pemberian Senyawa Trisindolina-5 Terhadap Ekspresi Gen c-Myc dan Apaf-1 Pada Kultur Sel Hela Secara In Vitro

Sulistyani, Rahma (2025) Studi Pengaruh Pemberian Senyawa Trisindolina-5 Terhadap Ekspresi Gen c-Myc dan Apaf-1 Pada Kultur Sel Hela Secara In Vitro. Other thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Ekspresi gen berperan penting dalam perkembangan kanker, seperti peningkatan gen proliferatif c-Myc dan penurunan gen pro-apoptosis APAF-1. Sel HeLa, berasal dari kanker serviks akibat integrasi HPV-18, digunakan untuk menguji efek Trisindolina-5. Trisindolina-5, senyawa alami golongan indole dan isatin dengan penambahan gugus fluor, diketahui memiliki aktivitas sitotoksik terhadap beberapa sel kanker seperti A549 dan DU-145 tanpa merusak sel normal, namun belum diuji terhadap sel HeLa. Penelitian ini bertujuan untuk mengevaluasi efek sitotoksik Trisindolina-5 terhadap sel HeLa dengan padanan obat Cisplatin sebagai kontrol positif menggunakan MTT assay, serta uji ekspresi gen APAF-1 dan c-Myc menggunakan qPCR untuk mengetahui aktivitas senyawa Trisindolina-5 pada jalur persinyalan apoptosis. Uji MTT assay menunjukkan IC50 Trisindolina-5 sebesar 297,42 µg/mL, lebih tinggi dibanding Cisplatin (3,78 µg/mL), menandakan efektivitas sitotoksik Cisplatin lebih kuat. Namun, analisis qPCR menunjukkan ekspresi c-Myc mengalami kenaikan dengan fold change 20.11 pada konsentrasi 100 µg/mL dan fold change 1.73 pada konsentrasi 200 µg/mL, hal ini berkesinambungan dengan ekspresi gen APAF-1 yang tidak mengalami peningkatan ekspresi yaitu fold change sebesar 0.812 pada konsentrasi 100 µg/mL dan fold change sebesar 0.057. Hasil ini diduga akibat sel HeLa mengalami nekrosis akibat stres oksidatif dan peningkatan ROS, bukan apoptosis intrinsik. Penelitian ini menunjukkan bahwa meskipun Trisindolina-5 memiliki aktivitas sitotoksik terhadap sel HeLa, mekanisme kematian sel yang terjadi masih perlu ditelusuri lebih lanjut.
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Gene expression plays a crucial role in cancer development, such as the upregulation of proliferative genes like c-Myc and the downregulation of pro-apoptotic genes like APAF-1. HeLa cells, derived from cervical cancer caused by HPV-18 integration, were used to examine the effect of Trisindoline-5. Trisindoline-5, a natural compound from the indole and isatin class with a fluorine substituent, has shown cytotoxic activity against cancer cell lines such as A549 and DU-145 without harming normal cells, but has not yet been tested on HeLa cells. This study aimed to evaluate the cytotoxic effect of Trisindoline-5 on HeLa cells, using Cisplatin as a positive control via MTT assay, and to assess APAF-1 and c-Myc gene expression using qPCR to understand the compound’s potential activity in the apoptosis signaling pathway. MTT assay results showed that the IC50 of Trisindoline-5 was 297.42 µg/mL, higher than Cisplatin (3.78 µg/mL), indicating that Cisplatin is more cytotoxic. However, qPCR analysis revealed that c-Myc expression did not significantly decrease and APAF-1 expression did not increase as hypothesized. c-Myc expression increased with a fold change of 20.11 at a concentration of 100 µg/mL and a fold change of 1.73 at a concentration of 200 µg/mL, this is consistent with the expression of the APAF-1 gene which did not experience an increase in expression, namely a fold change of 0.812 at a concentration of 100 µg/mL and a fold change of 0.057.This outcome is likely due to HeLa cells undergoing necrosis as a result of oxidative stress and elevated ROS, rather than intrinsic apoptosis. Another possibility is that apoptosis occurs via the extrinsic pathway, which does not involve APAF-1. This study indicates that while Trisindoline-5 exhibits cytotoxicity against HeLa cells, the underlying cell death mechanism requires further research.

Item Type:	Thesis (Other)
Uncontrolled Keywords:	APAF-1, c-Myc, Cisplatin, HeLa, Trisindolina-5
Subjects:	Q Science > QP Physiology > QP624 Molecular biology. Q Science > QR Microbiology > QR 201.T84 Tumors. Cancer
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Biology
Depositing User:	Rahma Sulistyani
Date Deposited:	29 Jul 2025 06:39
Last Modified:	29 Jul 2025 06:39
URI:	http://repository.its.ac.id/id/eprint/122612

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