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Journal : Makara Journal of Science

EKSPRESI RELATIF mRNA HIF-1α PADA JANTUNG, OTAK DAN DARAH TIKUS SELAMA INDUKSI HIPOKSIA SISTEMIK Wanandi, Septelia Inawati; Dewi, Syarifah; Paramita, Reni
Makara Journal of Science Vol. 13, No. 2
Publisher : UI Scholars Hub

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Abstract

Relative Expression of HIF-1α mRNA in Rat Heart, Brain and Blood During Induced Systemic Hypoxia. Hypoxia is a pathological condition in which the body as a whole or region of the body (tissue or cell) deprived of adequate oxygen supply. The transcriptional regulator hypoxia inducible factor-1 (HIF-1) is an essential mediator of O2 homeostasis. Unlike the β sub unit (HIF-1β), the activity of HIF-1α is controlled in an oxygen-dependent manner. It has been reported that the stability and expression of HIF-1α during hypoxia is remarkably higher than those under normoxic conditions.The aim of this study was to analyze the adaptive tissue responses during induced systemic hypoxia by comparation of relative expression of mRNA HIF-1α in rat heart, brain and blood. Twenty-five male Sprague Dawley rats were subjected to systemic hypoxia by placing them in the hypoxic chamber supplied by 8-10% of O2 for 0, 1, 7, 14 and 21 days, respectively. The relative expression level of HIF-1α mRNA in brain, heart and leucocyte cells were analyzed using quantitative RT-PCR assay (Real Time PCR) based on Pfaff’s formula. This study demonstrates that the increased of relative expression of HIF-1α mRNA during induced systemic hypoxia reached its maximum level at day 7 (in heart) or at day 14 (in brain), whereas in leucocyte cells the stimulation of HIF-1α expression was intensively maintained up to 21 days although the expression has reached the remarkably high level. We could conclude that HIF-1α as an oxygen sensing during systemic hypoxia has different capacity and sensitivity in brain, heart and blood tissues, due to the importance of oxygen homeostasis in each tissue
EKSPRESI RELATIF mRNA HIF-1α PADA JANTUNG, OTAK DAN DARAH TIKUS SELAMA INDUKSI HIPOKSIA SISTEMIK Wanandi, Septelia Inawati; Dewi, Syarifah; Paramita, Reni
Makara Journal of Science Vol. 13, No. 2
Publisher : UI Scholars Hub

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

Abstract

Relative Expression of HIF-1α mRNA in Rat Heart, Brain and Blood During Induced Systemic Hypoxia. Hypoxia is a pathological condition in which the body as a whole or region of the body (tissue or cell) deprived of adequate oxygen supply. The transcriptional regulator hypoxia inducible factor-1 (HIF-1) is an essential mediator of O2 homeostasis. Unlike the β sub unit (HIF-1β), the activity of HIF-1α is controlled in an oxygen-dependent manner. It has been reported that the stability and expression of HIF-1α during hypoxia is remarkably higher than those under normoxic conditions.The aim of this study was to analyze the adaptive tissue responses during induced systemic hypoxia by comparation of relative expression of mRNA HIF-1α in rat heart, brain and blood. Twenty-five male Sprague Dawley rats were subjected to systemic hypoxia by placing them in the hypoxic chamber supplied by 8-10% of O2 for 0, 1, 7, 14 and 21 days, respectively. The relative expression level of HIF-1α mRNA in brain, heart and leucocyte cells were analyzed using quantitative RT-PCR assay (Real Time PCR) based on Pfaff’s formula. This study demonstrates that the increased of relative expression of HIF-1α mRNA during induced systemic hypoxia reached its maximum level at day 7 (in heart) or at day 14 (in brain), whereas in leucocyte cells the stimulation of HIF-1α expression was intensively maintained up to 21 days although the expression has reached the remarkably high level. We could conclude that HIF-1α as an oxygen sensing during systemic hypoxia has different capacity and sensitivity in brain, heart and blood tissues, due to the importance of oxygen homeostasis in each tissue
MnSOD Gene Knockout Promotes Apoptosis in Triple-Negative Breast Cancer BT-549 Cells Through Survivin Inhibition and Caspase-3 and Caspase-9 Modulation Ghanny, Niken Rahmah; Wanandi, Septelia Inawati; Arumsari, Sekar; Paramita, Rafika Indah; Syahrani, Resda Akhra; Putri, Putu Indah Paramita Adi
Makara Journal of Science Vol. 28, No. 4
Publisher : UI Scholars Hub

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Abstract

Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype characterized by the lack of estro-gen, progesterone, and human epidermal growth factor receptor 2 (HER2) receptors. TNBC cells are becoming more aggressive due to the high expression of manganese superoxide dismutase (MnSOD) antioxidants to suppress reactive oxygen species-induced apoptosis and promote oncogenic signaling. This study was aimed to evaluate the effects of MnSOD knockout (KO) on TNBC cell apoptosis by assessing the survivin, caspase-9, and caspase-3 expressions. BT-549 TNBC cells containing the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-edited MnSOD gene were used to evaluate the messenger RNA (mRNA) expressions of survivin using the RT-qPCR assay. The West-ern blot assay was used to measure protein expressions of survivin, caspase-9, and caspase-3. Interactions between MnSOD and apoptosis-related proteins were simulated using computational methods based on molecular docking analysis and protein−protein interaction network. This study revealed that MnSOD KO decreased the binding affinity between MnSOD and survivin, in line with the significant reduction of survivin mRNA and protein expressions whereas the protein expressions of caspase-9 and caspase-3 increased in MnSOD KO cells. Therefore, MnSOD plays a pivotal role in BT-549 cell apoptosis by modulating survivin, caspase-9, and caspase-3 gene expressions. This study provides insights into a novel therapeutic strategy to mitigate the aggressiveness of TNBC by disrupting MnSOD gene expression. Further studies should elaborate the MnSOD signaling pathways involving closely related apoptotic proteins.
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 Vol. 29, No. 1
Publisher : UI Scholars Hub

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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