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All Journal ARTERI : Jurnal Ilmu Kesehatan Narra J Indonesian Journal of Cancer Makara Journal of Science Biosaintifika: Journal of Biology & Biology Education
Nihayah, Silviatun
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Electrical & Electronics Engineering Health Professions Immunology & microbiology Medicine & Pharmacology Nursing Public Health

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Pengembangan Metode PCR Multipleks untuk Analisis Genotipe Null Gen GSTM1/GSTT1 pada Pasien Tuberkulosis Prayuni, Kinasih; Razari, Intan; Nihayah, Silviatun; Syafrizal; Yuliwulandari, Rika
ARTERI : Jurnal Ilmu Kesehatan Vol 4 No 4 (2023): Agustus
Publisher : Puslitbang Sinergis Asa Professional

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37148/arteri.v4i4.289

Abstract

Tuberculosis (TB) remains Indonesia's leading infectious disease. Hepatotoxicity is the most common side effect of TB first-line medication therapy in TB patients. GSTM1 and GSTT1 are glutathione S-transferase (GST) genes involved in the detoxification of various toxic compounds such as drugs. The development of fast and simple methods for null genotyping of GSTM1/GSTT1 could facilitate large pharmacogenetic studies and the clinical application of personalized drug dose adjustment according to the patient's genetic profile. The aim of this research was to develop a multiple PCR method for simultaneous amplification of GSTM1/GSTT1 genes for molecular analysis. A total of 25 samples of TB patients were used to validate the method consisting of TB patients with hepatotoxicity and without hepatotoxicity. Our result showed the genotype frequency of the GSTM1 null genotype was 90% in TB patients with hepatotoxicity and 100% in TB patients without hepatotoxicity. The frequency of the GSTT1 null genotype in TB patients with hepatotoxicity was 90%, whereas in TB patients without hepatotoxicity was 80%. The sequencing results on the positive samples showed a similarity of 99% to the GenBank NCBI. Our study was successful in detecting GSTM1 and GSTT1 null genotypes using the multiplex PCR method in TB patients. Further study needs to be done with larger sample of TB patients.
Dual sgRNA-directed knock out survivin gene expression using CRISPR/Cas9 technology for editing survivin gene in triple-negative breast cancer Syahrani, Resda A.; Wanandi, Septelia I.; Arumsari, Sekar; Nihayah, Silviatun; Watanabe, Yukihide; Mizuno, Seiya; Louisa, Melva; Wuyung, Puspita E.
Narra J Vol. 4 No. 3 (2024): December 2024
Publisher : Narra Sains Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.52225/narra.v4i3.1177

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease 9 (CRISPR/Cas9) offers a robust approach for genome manipulation, particularly in cancer therapy. Given its high expression in triple-negative breast cancer (TNBC), targeting survivin with CRISPR/Cas9 holds promise as a therapeutic strategy. The aim of this study was to design specific single guide ribonucleic acid (sgRNA) for CRISPR/Cas9 to permanently knock out the survivin gene, exploring its potential as a therapeutic approach in breast cancer while addressing potential off-target effects. Survivin gene knockout was conducted in the TNBC cell line BT549. Intron 1, exon 2, and intron 2 of the survivin gene were selected as sgRNA targets. These sgRNAs were designed in silico and then cloned into a CRISPR/Cas9 expression plasmid. The cleavage activity was assessed using an enhanced green fluorescent protein (EGFP) expression plasmid. The sgRNAs with higher cleavage activity were selected for the establishment of knockout cells. After transfecting the plasmid into the cells, the success of the survivin gene knockout was validated at the deoxyribonucleic acid (DNA) level using polymerase chain reaction (PCR) and sequencing analysis, and at the protein expression level using Western blotting. The study found that sgRNAs survin1A (targeting intron 1), survex2A (targeting intron 2), and survin2A (targeting intron 2) demonstrated higher cleavage activities compared to the other sgRNAs. However, using the single sgRNA, survex2A did not generate mutations in the survivin gene. At the protein level, survivin was still expressed, indicating that a single sgRNA was ineffective in knocking out the survivin gene. In contrast, the combination of sgRNA survin1A and sgRNA survin2A was more effective in generating mutations in the survivin gene, resulting in the deletion of the entire exon 2 and leading to a loss of survivin protein expression. In conclusion, our work provides specific sgRNAs and demonstrates the utilization of dual sgRNAs strategy in the CRISPR/Cas9 technology to knock out the survivin gene, showing potential in breast cancer therapy.
Regulation of Survivin Gene Expression in Stem Cell and Cancer Stem Cells and Emphasis Approach Nihayah, Silviatun; Wanandi, Septelia Inawati
Indonesian Journal of Cancer Vol 19, No 1 (2025): March
Publisher : http://dharmais.co.id/

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33371/ijoc.v19i1.1142

Abstract

Background: Gene expression regulation is a method that cells utilize to enhance or decrease the output of specific genes (proteins or RNA). In biological and medical research such as cancer, gene expression is routinely observed. Survivin is a protein that is commonly produced in cancer cells and has the potential to be investigated. Survivin is a unique protein with two distinct roles: preventing apoptosis and regulating cell division. The inhibitory apoptosis protein (IAP) family includes the smallest member. Furthermore, Survivin is highly expressed in a variety of somatic stem cell types as well as human embryonic stem cells. Comparing most cancer cells to normal tissues, survivin is also present in higher amounts. This review aimed to examine the properties of survivin, how it is expressed in cancerous and normal stem cells, how it affects proliferation or apoptosis, and how to block their expression. Methods: The literature on the regulation of survivin expression in cancer cells and stem cells that was published in English between 2014 and 2024 is reviewed in this study. For articles, we looked through PubMed, Scopus, and the Google Scholar database. In order to support the theories, publications from before 2014 were also tracked down.Results: Molecular mechanism studies indicate that survivin participates in numerous signaling pathways, including MAPK, STAT3, b-catenin, Wnt, Notch, and others, and also controls the progression of the cell cycle and cytokinesis. Several elements, such as signaling pathway blockage siRNA technology, and CRISPR/Cas9 system have been discovered to aid in the induction of cancer cell death. Conclusion: Survivin is linked to several cancer survival-related pathways, contributing to carcinogenesis. Its expression is associated with treatment resistance, tumor development, and poor prognosis.
Roles of the Survivin BIR Domain in Cellular Apoptosis and Proliferation: An In Silico Study Nihayah, Silviatun; Wanandi, Septelia Inawati; Erlina, Linda; Syahrani, Resda Akhra
Makara Journal of Science
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Survivin is an antiapoptotic protein that is highly expressed in cancer cells. We investigated the dual roles of the Bacu-lovirus IAP Repeats (BIR) domain within survivin, encompassing both apoptosis and proliferation, through an in silico study. The protein-protein interaction (PPI) network of survivin was analyzed using Cytoscape software. Functional enrichment (FE) analysis and data from the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to identify the implicated signaling pathways. The binding affinity of the BIR domain with the targeted proteins was visualized via molecular docking analysis. Drawing insights from the PPI network and FE analysis, we identified two key proteins in-volved in apoptosis such as X-linked Inhibitor of Apoptosis Proteins (XIAP) and caspase-9, and proliferation such as Cyclin-dependent Kinase 1 (CDK1) and Inner Centromere Protein (INCENP) for further analysis of their binding with the survivin BIR domain. These proteins were found to bind to the BIR domain at the Thr34, Thr48, and Ser20 resi-dues that have critical roles to regulate the apoptosis and proliferation. This study provides future insights into how the BIR domain of survivin could emerge as a potential target for cancer treatment, such as determining knockout targets for the development of genome editing technology
Correlation of Human Telomerase Reverse Transcriptase Promoter (hTERT) Gene Methylation and Ageing Nihayah, Silviatun; Wening Sari; Yusnita, Yusnita; Intan Razari; Kinasih Prayuni; Utomo, Ahmad Rusdan Handoyo
Biosaintifika: Journal of Biology & Biology Education Vol. 17 No. 2 (2025): August 2025
Publisher : Universitas Negeri Semarang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/biosaintifika.v17i2.22038

Abstract

Promoter methylation of the hTERT gene in blood DNA has been proposed to be an epigenetic molecular clock because it is negatively correlated with ageing. Saliva is an alternative source of DNA because it contains buccal cells. Currently, the correlation of hTERT promoter methylation and ageing using saliva is not known. This study aimed to determine: first, the correlation between hTERT promoter methylation and ageing in saliva, as a source of non-invasive DNA sampling, instead of blood was determined. Second, the influence of sex on the methylation of the hTERT promoter in ageing was evaluated. A cross-sectional study design was used, and 119 subjects were recruited, consisting of 25 children (1-5 y.o), 42 teens (17-19 y.o), 16 adults (20-50 y.o), and 36 elderly (60 to 84 y.o). Promoter methylation of the hTERT of extracted DNA was determined using the MSRE (methyl-specific restriction enzyme) method. The relationship between age and the percentage of hTERT methylation was assessed using the Pearson test. The percentage of hTERT methylation in saliva DNA was negatively correlated with age r= -0.4305 (p-value <0.05). Negative correlation was also found in men (r= -0.376) and women (r=-0.43). Negative correlation between hTERT and ageing has been confirmed in saliva as a noninvasive sampling method. The benefit of this research in ethical and social considerations may encourage a greater participation in ageing research, particularly in underprivileged communities, thus democratising access to scientific advancements in longevity studies.
Co-Authors Erlina, Linda Intan Razari Kinasih Prayuni Kinasih Prayuni, Kinasih Melva Louisa Mizuno, Seiya Puspita E. Wuyung Razari, Intan Rika Yuliwulandari, Rika SEKAR ARUMSARI, SEKAR Septelia I. Wanandi SEPTELIA INAWATI WANANDI syafrizal Syahrani, Resda A. Syahrani, Resda Akhra Utomo, Ahmad Rusdan Handoyo Watanabe, Yukihide Wening Sari YUSNITA YUSNITA