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

• Published in: Cytotechnology (Dordrecht) Vol. 68; no. 6; pp. 2311 - 2321
• Main Authors: Lavanya, Ch, Sibin, M. K., Srinivas Bharath, M. M., Manoj, M. Jeru, Venkataswamy, Manjunatha M., Bhat, Dhananjaya I., Narasinga Rao, K. V. L., Chetan, G. K.
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01-12-2016; Springer Nature B.V
• Subjects: Apoptosis; Biochemistry; Biomarkers; Biomedicine; Biotechnology; Brain cancer; Brain tumors; Carcinogenesis; Catalytic subunits; Cell death; Cell viability; Chemistry; Chemistry and Materials Science; Chromosomes; Down-regulation; Flow cytometry; Gene expression; Glioblastoma; Glioma; Immunoblotting; Laboratories; Medical prognosis; Original; Original Article; Patients; RNA polymerase; RNA-directed DNA polymerase; RNA-mediated interference; siRNA; Stem cells; Telomerase; Telomerase reverse transcriptase; Tumors; Yeast; United States > US; India; Small interfering RNA; LN18 cell line; hTERT; Glioblastoma; Apoptosis
• ISSN: 0920-9069; 1573-0778
• DOI: 10.1007/s10616-016-0025-8

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.