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1. |
MULTIPLE GENE TARGETING SIRNAS FOR DOWN REGULATION OF IMMEDIATE EARLY-2 (IE2) AND DNA POLYMERASE GENES MEDIATED INHIBITION OF NOVEL RAT CYTOMEGALOVIRUS (STRAIN ALL-03)
DESCRIPTION : Cytomegalovirus (CMV) is an opportunistic pathogen that causes severe complications in congenitally infected newborns and non-immunocompetent individuals. Developing an effective vaccine is a major public health priority and current drugs are fronting resistance and side effects on recipients. In the present study, with the aim of exploring new strategies to counteract CMV replication, several anti-CMV siRNAs targeting IE2 and DNA polymerase gene regions were characterized and used as in combinations for antiviral therapy. Methods The rat embryo fibroblast (REF) cells were transfected with multi siRNA before infecting with CMV strain ALL-03. Viral growth inhibition was measured by tissue culture infectious dose (TCID50), cytopathic effect (CPE) and droplet digital PCR (ddPCR) while IE2 and DNA polymerase gene knockdown was determined by real-time PCR. Ganciclovir was deployed as a control to benchmark the efficacy of antiviral activities of respective individual siRNAs. Results There was no significant cytotoxicity encountered for all the combinations of siRNAs on REF cells analyzed by MTT colorimetric assay (P > 0.05). Cytopathic effects (CPE) in cells infected by RCMV ALL-03 had developed significantly less and at much slower rate compared to control group. The expression of targeted genes was downregulated successfully resulted in significant reduction (P < 0.05) of viral mRNA and DNA copies (dpb + dpc: 79%, 68%; dpb + ie2b: 68%, 60%; dpb + dpc + ie2b: 48%, 42%). Flow cytometry analysis showed a greater percentage of viable and early apoptosis of combined siRNAs-treated cells compared to control group. Notably, the siRNAs targeting gene regions were sequenced and mutations were not encountered, thereby avoiding the formation of mutant with potential resistant viruses. Conclusions In conclusion. The study demonstrated a tremendous promise of innovative approach with the deployment of combined siRNAs targeting at several genes simultaneously with the aim to control CMV replication in host cells. CONTRIBUTOR : MOHD AZMI BIN MOHD LILA (11/05/2023 15:43 PM)
FACULTY : FAKULTI PERUBATAN VETERINAR
PUBLICATION : 0 ; FILE IN : HDFS HAS VIEWS : 4872 ; FOLDER IN PROJECT : 1 ; FILE IN PROJECT : 1 ; DOWNLOAD : 0 ; KEYWORD : [Cytomegalovirus] , [ Combinations] , [ Small interfering RNA] , [ Replication] , [ Gene expression] |
2. |
HUMAN MESENCHYMAL STEM CELLS-MEDIATED TRANSCRIPTOMIC REGULATION OF LEUKEMIC CELLS IN DELIVERING ANTI-TUMORIGENIC EFFECTS
DESCRIPTION : Treatment of leukemia has become much difficult because of resistance to the existing anticancer therapies. This has thus expedited the search for alternativ therapies, and one of these is the exploitation of mesenchymal stem cells (MSCs) towards control of tumor cells. The present study investigated the effect of human umbilical cord-derived MSCs (UC-MSCs) on the proliferation of leukemic cells and gauged the transcriptomic modulation and the signaling pathways potentially affected by UC-MSCs. The inhibition of growth of leukemic tumor cell lines was assessed by proliferation assays, apoptosis and cell cycle analysis. BV173 and HL-60 cells were further analyzed using microarray gene expression profiling. The microarray results were validated by RT-qPCR and western blot assay for the corresponding expression of genes and proteins. The UC-MSCs attenuated leukemic cell viability and proliferation in a dose-dependent manner without inducing apoptosis. Cell cycle analysis revealed that the growth of tumor cells was arrested at the G0/G1 phase. The microarray results identified that HL-60 and BV173 share 35 differentially expressed genes (DEGs) (same expression direction) in the presence of UC-MSCs. In silico analysis of these selected DEGs indicated a significant influence in the cell cycle and cell cycle-related biological processes and signaling pathways. Among these, the expression of DBF4, MDM2, CCNE2, CDK6, CDKN1A, and CDKN2A was implicated in six different signaling pathways that play a pivotal role in the anti-tumorigenic activity exerted by UC-MSCs. The UC-MSCs perturbate the cell cycle process of leukemic cells via dysregulation of tumor suppressor and oncogene expression. CONTRIBUTOR : LING KING HWA (11/05/2023 14:43 PM)
FACULTY : FAKULTI PERUBATAN DAN SAINS KESIHATAN
PUBLICATION : 0 ; FILE IN : HDFS HAS VIEWS : 3129 ; FOLDER IN PROJECT : 1 ; FILE IN PROJECT : 1 ; DOWNLOAD : 0 ; KEYWORD : [cell cycle; gene expression profiling; leukemia; mesenchymal stem cells] |
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