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RNA interference mediated downregulation of human telomerase reverse transcriptase (hTERT) in LN18 cells

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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.

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Acknowledgments

Lavanya Ch was supported by a UGC RGNF-Senior research fellowship (University Grants Commission- Rajiv Gandhi National Fellowship), New Delhi, India. The study was financially supported by DST-SERB (Department of Science and Technology-Science and Engineering Research Board), Government of India. We thank Dr. Pandareesh MD for technical help and data analysis. We thank Dr. Hemanth, Dr. Anu Mary Vargeesh for helping in cell culture work.

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Authors and Affiliations

  1. Department of Human Genetics, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India

    Ch. Lavanya, M. K. Sibin, M. Jeru Manoj & G. K. Chetan

  2. Department of Neuro-chemistry, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India

    M. M. Srinivas Bharath

  3. Department of Neurovirology, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India

    Manjunatha M. Venkataswamy & K. V. L. Narasinga Rao

  4. Department of Neurosurgery, National Institute of Mental Health and Neuro Sciences, Bangalore, India

    Dhananjaya I. Bhat

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  1. Ch. Lavanya
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Correspondence to G. K. Chetan.

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Lavanya, C., Sibin, M.K., Srinivas Bharath, M.M. et al. RNA interference mediated downregulation of human telomerase reverse transcriptase (hTERT) in LN18 cells. Cytotechnology 68, 2311–2321 (2016). https://doi.org/10.1007/s10616-016-0025-8

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