Silencing of the hTERT Gene by shRNA Inhibits Colon Cancer SW480 Cell Growth In Vitro and In Vivo

Abstract

Human telomerase reverse transcriptase (hTERT) is the key enzyme responsible for synthesizing and maintaining the telomeres on the ends of chromosomes, and it is essential for cell proliferation. This has made hTERT a focus of oncology research and an attractive target for anticancer drug development. In this study, we designed a small interfering RNA (siRNA) targeting the catalytic subunit of hTERT and tested its effects on the growth of telomerase-positive human colon carcinoma SW480 cells in vitro, as well as on the tumorigenicity of these cells in nude mice. Transient and stable transfection of hTERT siRNA into colon cancer SW480 cells suppressed hTERT expression, reduced telomerase activity and inhibited cell growth and proliferation. Knocking down hTERT expression in SW480 tumors xenografted into nude mice significantly slowed tumor growth and promoted tumor cell apoptosis. Our results suggest that hTERT is involved in carcinogenesis of human colon carcinoma, and they highlight the therapeutic potential of a hTERT knock-down approach.

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

“…Knock-down of hTERT by siRNA significantly decreases hTERT expression and induces apoptosis through activation of multiple pathways viz., mitochondrial signal transduction pathway, PI3 K/AKT pathway, Bax/Bcl-2 (Wang et al 2007(Wang et al , 2012Shi et al 2014). Both transient and stable transfection of hTERT siRNA in SW480 colon cancer cells inhibits hTERT expression, down regulates telomerase activity and suppresses cell proliferation (Liu et al 2014). …”

CD13: A Key Player in Multidrug Resistance in Cancer Chemotherapy

“…This suggests that MRP2 may be involved in the development of intrinsic MDR in HCC, and MRP5 and MRP3 expression may be related to acquired MDR in HCC. CD13 expression was positively correlated with the expression of MRP2 and MRP3 in HCC cells and LCSCs, indicating that CD13 mediates intrinsic and extrinsic MDR in HCC cells by increasing drug efflux 56 . MRPs belong to the ABC superfamily along with P-gp, and MRPs can pump drugs with the help of ATP-dependent glutathione-binding S carrier (GS-X pump), for which the activity of glutathione-S-transferases (GSTs), including GST-π, GST-α, and GST-μ, was essential 57 .…”

Ferritin-Nanocaged ATP Traverses the Blood–Testis Barrier and Enhances Sperm Motility in an Asthenozoospermia Model

“…The HFR shRNA sequences were synthesized commercially (Tsingke) (Table S4) and cloned into vector pLKO.1-mcherry-puro (MiaoLing, P10494, Wuhan, China) using the protocol described elsewhere. 54 The HEK-293FT packaging cells were transfected with the pLKO.1-shHFR plasmid and the packaging plasmids including pRSV-Rev (addgene ID:12253), pCMV-VSV-G (addgene ID:8454), and pMDLg/pRRE (addgene ID:12251). After 10 h of transfection, the cultured medium was replaced with a fresh medium.…”