Studi Pengaruh Senyawa Trisindolina 1 Terhadap Ekspresi Gen P53 Dan P53R2 pada Cancer Stem Cell (CSC)

Ghaissani, Shabrina Syifa (2022) Studi Pengaruh Senyawa Trisindolina 1 Terhadap Ekspresi Gen P53 Dan P53R2 pada Cancer Stem Cell (CSC). Masters thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Cancer Stem Cell (CSC) memiliki karakteristik stemness yaitu mampu melakukan self-renewal dan diferensiasi yang dipengaruhi oleh faktor transkripsi, tumor-specific microenvironment dan signaling protein. CSC berperan penting dalam perkembangan tumor, rekurensi, dan resistansi terhadap kemoterapi maka potensi CSC sebagai target terapi sangat besar. Salah satu faktor penyebab terjadinya DNA repair pada CSC adalah ekspresi protein P53R2 yang diketahui berkorelasi dengan progonosis buruk pada pasien kanker. Agen perusak DNA menginduksi ekspresi p53R2 dengan cara yang bergantung pada p53 dan p53R2. Protein P53R2 merupakan building block untuk replikasi dan perbaikan DNA. Protein tumor suppressor P53 juga mengontrol self- renewal dan diferensiasi. Tujuan dari penelitian ini untuk mengetahui dan menganalisis senyawa Trisindolina 1 pada regulasi gen yang berperan dalam stemness CSC yaitu gen P53R2 dan P53. Metode yang dilakukan adalah uji sitotoksisitas dengan metode MTT assay. Sel yang digunakan CS. Kosentrasi yang digunakan 1; 2,5; 5; 10; 25; 50 dan 100 μg/mL dengan 3 ulangan. Doxorubicin sebagai control. Uji apoptosis dengan metode flow cytometry dengan konsentrasi 1/8 IC50, ¼ IC50, ½ IC50 dan 1 IC50 inkubasi 24h. Analisis data apoptosis dilakukan dengan aplikasi Cellquest. Uji Ekspresi gen P53R2 digunakan Real Time PCR. Ekspresi gen secara kuantitatif pra dan post perlakuan Trisindolina 1 digunakan CT Value. Data qRT-PCR dianalisis menggunakan Metode Livak. Hasil uji sitotoksisitas Trisindolina 1 diperoleh nilai IC50 sebesar 60 μg/mL. Uji Apotosis tertinggi diperoleh pada sel dengan perlakuan doxorubicin 10 μg/mL dengan persentase 98,2 0,88 % sedangkan persentase apoptosis tertinggi oleh perlakuan Trisindolina ditemukan pada perlakuan IC50 dengan nilai 12,6 0,96 %. Ekspresi gen P53 tertinggi maupun ekspresi P53R2 terendah ditemukan pada sel dengan perlakuan Trisindolina IC50 dengan nilai sedangkan peningkatan ekspresi masing masing berurutan adalah 33,13 dan 1,57 kali lipat.
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Cancer Stem Cells (CSC) have stemness characteristics that are capable of self-renewal and differentiation which is influenced by transcription factors, tumorspecific microenvironment and signaling proteins. CSCs play an important role in
tumor development, recurrence, and resistance to chemotherapy, so the potential for CSCs as therapeutic targets is very large. One of the factors causing DNA repair in CSC is the expression of the P53R2 protein which is known to correlate with poor prognosis in cancer patients. DNA-damaging agents induce p53R2 expression in a p53 and p53R2-dependent manner. P53R2 protein is a building block for DNA replication and repair. The tumor suppressor protein P53 also controls self-renewal and differentiation. The purpose of this study was to determine and analyze the Trisindolina 1 compound in the regulation of genes that play a role in CSC stemness, namely the P53R2 and P53 genes. The method used is the cytotoxicity test using the MTT assay method. Cells used by CS. The concentration used is 1; 2.5; 5; 10; 25; 50 and 100 g/mL with 3 replicates. Doxorubicin as a control. Apoptosis test using flow cytometry method with a concentration of 1/8 IC50, IC50, IC50 and 1 IC50 incubation 24h. Apoptotic data analysis was performed using the Cellquest application. P53R2 gene expression test used Real Time PCR. Gene expression quantitatively before and after Trisindolina 1 treatment used CT Value. The qRT-PCR data were analyzed using the Livak Method. The results of the Trisindolina 1 cytotoxicity test obtained an IC50 value of 60 g/mL. The highest apoptosis test was obtained in cells treated with doxorubicin 10 g/mL with a percentage of 98.2 0.88%, while the highest percentage of apoptosis by Trisindolina treatment was found in IC50 treatment with a value of 12.6 0.96%. The highest P53 gene expression and the lowest P53R2 expression were found in cells treated with Trisindolina IC50 with a value of 33.13 and 1.57 times the increase in expression, respectively.

Item Type:	Thesis (Masters)
Uncontrolled Keywords:	Ekspresi Gen, Apoptosis, P53R2, P53, CSC, Trisindolina 1, Gene Expression, Apoptosis
Subjects:	Q Science > QR Microbiology > QR 201.T84 Tumors. Cancer
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Biology > 46101-(S2) Master Thesis
Depositing User:	Mr. Marsudiyana -
Date Deposited:	15 Dec 2025 01:48
Last Modified:	15 Dec 2025 01:48
URI:	http://repository.its.ac.id/id/eprint/128959

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