title: RNA interference mediated downregulation of human telomerase reverse transcriptase (hTERT) in LN18 cells.
abstract: Human telomerase reverse transcriptase (hTERT) gene is a biomarker for the targeted therapy in various cancers. Presence of increased telomerase activity is a common feature of all cancers including glioblastoma. Both RNA and catalytic subunits of hTERT are the target sites for blocking its activity. The current study focuses on the expression of hTERT in glioblastoma and its regulation using two different novel siRNAs (small interfering RNA). Our patient data demonstrated increased expression of hTERT, which could be correlated with carcinogenesis in glioma. In vitro studies in siRNA transfected LN18 cells confirmed significant cell death (p < 0.05) as evidenced by MTT and trypan blue exclusion assay. These results were further supported by flow cytometry data, which showed significant increase in early and late apoptosis. The hTERT mRNA expression was effectively downregulated by 45 and 39 % with siRNA1 and siRNA2, respectively. These results were further confirmed by immunoblotting analysis (p < 0.05). Our results suggest that both the siRNAs effectively down regulated the expression of hTERT at mRNA and protein levels, thereby decreasing cell viability and proliferation rate. Hence siRNA mediated downregulation of hTERT could be a potential therapeutic avenue in glioblastoma.
authors.affiliation: Department of Human Genetics, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India.
authors.forename: Ch
authors.initials: C
authors.lastname: Lavanya
authors.affiliation: Department of Human Genetics, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India.
authors.forename: M K
authors.initials: MK
authors.lastname: Sibin
authors.affiliation: Department of Neuro-chemistry, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India.
authors.forename: M M
authors.initials: MM
authors.lastname: Srinivas Bharath
authors.affiliation: Department of Human Genetics, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India.
authors.forename: M Jeru
authors.initials: MJ
authors.lastname: Manoj
authors.affiliation: Department of Neurovirology, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India.
authors.forename: Manjunatha M
authors.initials: MM
authors.lastname: Venkataswamy
authors.affiliation: Department of Neurosurgery, National Institute of Mental Health and Neuro Sciences, Bangalore, India.
authors.forename: Dhananjaya I
authors.initials: DI
authors.lastname: Bhat
authors.affiliation: Department of Neurovirology, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India.
authors.forename: K V L
authors.initials: KV
authors.lastname: Narasinga Rao
authors.affiliation: Department of Human Genetics, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India. drchetangk@gmail.com.
authors.forename: G K
authors.initials: GK
authors.lastname: Chetan
keywords: Apoptosis; Glioblastoma; LN18 cell line; Small interfering RNA; hTERT