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The intrabody targeting of hTERT attenuates the immortality of cancer cells

Abstract

hTERT (human telomerase reverse transcriptase) plays a key role in the process of cell immortalization. Overexpression of hTERT has been implicated in 85% of malignant tumors and offers a specific target for cancer therapy. In this paper, we describe an effective approach using a single-chain variable fragment (scFv) intrabody derived from monoclonal hybridoma directed against hTERT to attenuate the immortalization of human uterine cervix and hepatoma cells. The scFv we constructed had a high affinity to hTERT, and specifically neutralized over 70% of telomere synthesis activity, thereby inhibiting the viability and proliferation of the cancer cells. Our results indicate that this anti-hTERT intrabody is a promising tool to target hTERT and intervene in the immortalization process of cancer cells.

Abbreviations

DAPI:

diamidino-phenyl-indole

ECL:

enhanced chemiluminescence

ELISA:

enzyme linked immunosorbent assay

FITC:

fluorescein isothiocyanate

HRP:

horseradish peroxidase

mAb:

monoclonal antibody

MIFs:

mouse intraperitoneal fluid

MTT:

monotetrazolium

PI:

propidium iodide

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Correspondence to Daming Ren.

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Zhu, X., Yang, N., Cai, J. et al. The intrabody targeting of hTERT attenuates the immortality of cancer cells. Cell Mol Biol Lett 15, 32 (2010). https://doi.org/10.2478/s11658-009-0032-2

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  • DOI: https://doi.org/10.2478/s11658-009-0032-2

Key words

  • Cancer
  • Intrabody
  • Anti-hTERT ScFv
  • Immortality