Effect of berberine on HCT 116 colon carcinoma cell proliferation, telomerase activities, and next generation sequencing transcriptome data profiling / Nadhirah Ahmad

Nadhirah , Ahmad (2021) Effect of berberine on HCT 116 colon carcinoma cell proliferation, telomerase activities, and next generation sequencing transcriptome data profiling / Nadhirah Ahmad. Masters thesis, Universiti Malaya.

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Abstract

Telomerase is an RNA-dependent DNA polymerase that synthesizes telomeric DNA repeats (TTAGGG) at the 3’ end of chromosomes. Telomerase is gaining attention as a prospective molecular target in anticancer therapies as it is found to be highly expressed in most cancerous cells but scarcely detectable in normal somatic cells. Although inhibiting telomerase activity could be a promising strategy to combat cancer, the transcriptome changes upon telomerase inhibition is not well documented. In this study, human tumor cell lines were treated with telomerase inhibitors and next generation sequencing-based transcriptomic profiling analysis were conducted. First, the effect of telomerase inhibitors i.e., silymarin, boldine, and berberine on selected cancer cell lines’ (HCT 116, A549 and CaSki) cell proliferation were evaluated using SRB assay. Berberine exerted the highest cytotoxic activity against HCT 116 cells in a time and dose-dependent manner in comparison to A549 and CaSki cell lines. Subsequent telomerase inhibition analysis was conducted at 8 μg/ml of berberine, 24 hours treatment on HCT 116 cell lines. Berberine exposure to HCT 116 cells resulted in the downregulation of telomerase related genes expression which were detected via Western blot and Real-Time PCR. RNA-Seq analysis revealed a total of 21,179 and 19,793 genes identified from the control and berberine treated datasets, respectively. Comparative analysis revealed a total of 324 differentially expressed genes (DEGs) with 48 upregulated genes and 276 downregulated genes. The DEGs were subjected to gene annotation analysis and were predicted to be involved in the regulation of protein kinases, response to endoplasmic reticulum stress, cellular response to glucose starvation, RNA metabolic processes, and cell death. All these results shed light to the potential use of berberine as anti-telomerase-based therapy.

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