Pengaruh mimic hsa-Mir-203a-3p Terenkapsulasi Eksosom terhadap Viabilitas dan Migrasi Sel Hs578T: Validasi Eksperimental dan Prediksi Gen Target secara In Silico

Syamsul Arif Ardiansyah, Prof. Sofia Mubarika Haryana M.Med. Sc., Ph.D

2024 | Tesis | S2 Bioteknologi

Triple-negative breast cancer (TNBC) merupakan jenis kanker yang sangat berbahaya karena sifatnya yang heterogen, invasif, dan resisten terhadap terapi. Salah satu strategi potensial adalah penggunaan miRNA, khususnya Hsa-miR-203a-3p, yang diketahui mengalami penurunan regulasi pada TNBC. Dalam penelitian ini, digunakan mimic-Hsa-miR-203a-3p sebagai tiruan hsa-miR-203a-3p endogen. Namun, karena sifat miRNA yang tidak stabil, diperlukan agen pengantar seperti eksosom yang stabil, tidak toksik, dan non-imunogenik sebagai agen enkapsulasi untuk miRNA, membentuk eksomiR. Tujuan penelitian ini adalah untuk mengevaluasi karakteristik eksosom yang digunakan, yang berasal dari sekretom Umbilical cord mesenchymal stem cell (UCMSC), serta pengaruh eksomiR terhadap viabilitas dan migrasi sel TNBC Hs578T. Selain itu, penelitian ini juga bertujuan untuk mengamati ekspresi hsa-miR-203a-3p setelah penambahan eksomiR dan mengidentifikasi gen target hsa-miR-203a-3p melalui analisis in silico. Viabilitas sel diuji menggunakan metode MTT assay, sementara migrasi sel dianalisis menggunakan wound healing assay. Ekspresi gen dievaluasi dengan qRT-PCR. Analisis in silico dilakukan menggunakan dataset dari TargetScan, TarBase, dan GSEA, serta analisis metabolisme dilakukan dengan David dan Panther. Analisis prognosis menggunakan PrognoScan dan visualisasi data dilakukan dengan Cytoscape. Hasil penelitian menunjukkan bahwa pada volume 2x, viabilitas sel tehambat hingga 53?ngan signifikansi P<0>

Triple-negative breast cancer (TNBC) is a highly dangerous type of cancer due to its heterogeneous, invasive, and therapy-resistant nature. One potential strategy is the use of miRNAs, specifically Hsa-miR-203a-3p, which is known to be downregulated in TNBC. In this study, mimic-Hsa-miR-203a-3p was used to replicate the endogenous hsa-miR-203a-3p. However, due to the unstable nature of miRNA, a delivery agent such as exosomes is required. Exosomes are stable, non-toxic, and non-immunogenic, making them suitable encapsulation agents for miRNA, forming exomiR. The aim of this study is to evaluate the characteristics of the exosomes used, which are derived from the secretome of human umbilical cord mesenchymal stem cells (UCMSC), and to assess the effect of exomiR on the viability and migration of TNBC Hs578T cells. Additionally, the study aims to observe the expression of hsa-miR-203a-3p after the addition of exomiR and to identify the target genes of hsa-miR-203a-3p through in silico analysis. Cell viability was tested using the MTT assay, while cell migration was analyzed using the wound healing assay. Gene expression was evaluated by qRT-PCR. In silico analysis was conducted using datasets from TargetScan, TarBase, and GSEA, and metabolism analysis was performed using DAVID and Panther. Prognosis analysis was conducted using PrognoScan, and data visualization was done with Cytoscape. The results showed that at 2x volume, cell viability was inhibited by up to 53% with a significance of P<0>

Kata Kunci : Triple Negative Breast Cancer, EksomiR, Viabilitas, Migrasi, Ekspresi miRNA