Zulkifli, Nur Diyana (2025) Global Differential Gene Expression In Hypoxic Hepg2 Upon Sirtuin-1 Upregulation And Downregulation. Masters thesis, Perpustakaan Hamzah Sendut.
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Abstract
Hepatocellular carcinoma (hcc) is one of the leading causes of cancer death worldwide, largely due to its poor prognosis and the emergence of chemoresistance, particularly exacerbated by the hypoxic tumor microenvironment (tme). Sorafenib, one of the targeted cancer drugs, is commonly used to treat advanced hcc. However, this treatment frequently induces drug resistance. Sirtuin-1 (sirt1) is found to be associated with chemoresistance and linked to tumor angiogenesis under hypoxia, which may serve as a potential molecular target for hcc treatment. Sirt1 is significantly dysregulated in various cancers, including hcc. Therefore, this study aims to elucidate sirt1 as a potential molecular target for hcc progression in hypoxic conditions. To achieve this objective, crispr/dcas9 was utilized to modulate sirt1 gene expression in hepg2 cells. 20 bp grnas targeting sirt1 were cloned into the plasmid vector pspgrna and verified by sequencing. Sirt1 upregulation and downregulation were accomplished by transducing hepg2 cells with vp64-activator and krab-repressor plasmids, followed by antibiotic selection to establish hepg2_crispra_vp64 and hepg2_crispri_krab stable cells. The transfection with grna was done in normoxic condition followed by incubation in hypoxia. Result on optimization shows upregulation and downregulation to 1.7- and 0.4-fold change in transfected hepg2_crispra_vp64 and hepg2_crispri_krab stable cells, respectively.
| Item Type: | Thesis (Masters) |
|---|---|
| Subjects: | R Medicine > R Medicine (General) > R5-920 Medicine (General) |
| Divisions: | Institut Perubatan & Pergigian Termaju (Advanced Medical & Dental Institute (AMDI)) > Thesis |
| Depositing User: | Mr Hasmizar Mansor |
| Date Deposited: | 04 Jun 2026 07:02 |
| Last Modified: | 04 Jun 2026 07:02 |
| URI: | http://eprints.usm.my/id/eprint/64295 |
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