@article{APS3785,
author = {Sheng-lin Huang and Yi Wu and Hai Yu and Ping Zhang and Xing-qian Zhang and Lei Ying and Han-fang Zhao},
title = {Inhibition of Bcl-2 expression by a novel tumor-specific RNA interference system increases chemosensitivity to 5-fluorouracil in Hela cells},
journal = {Acta Pharmacologica Sinica},
volume = {27},
number = {2},
year = {2016},
keywords = {},
abstract = {Aim: RNA interference (RNAi) has been proposed as a potential treatment for
cancer, but the lack of cellular targets limits its use in cancer gene therapy. No
current technology has achieved direct tumor-specific gene silencing using RNAi.
In the present study we attempt to develop a tumor-specific RNAi system using
the human telomerase reverse transcriptase (hTERT) promoter; furthermore, we
analyzed its inhibitive effect on Bcl-2 expression. Methods: The vectors containing
a small hairpin RNA (shRNA) to target exogenous reporters [firefly luciferase
and enhanced green fluorescent protein (EGFP)] and endogenous gene (Bcl-2)
were constructed. Luciferase expression was determined by dual luciferase assay.
Reverse transcription-polymerase chain reaction (RT-PCR), fluorescence microscopy
and fluorescence-activated cell sorting (FACS) were used to measure EGFP
expression. Inhibition of Bcl-2 was evaluated by RT-PCR and Western blotting.
Cell proliferation and viability were measured by using 3-(4,5-dimethylthiazol-2-
yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. FACS was used to analyze
the cell cycle distribution profile. Results: We showed that with the hTERT
promoter directly driving shRNA transcription, expression of the exogenous reporters
(LUC and EGFP) in tumor cells, but not normal cells, was specifically
inhibited in vitro. The hTERT promoter-driven shRNA also depressed the expression
of Bcl-2. Inhibition of Bcl-2 did not affect cell proliferation, but increased the
chemosensitivity of HeLa cells to 5-fluorouracil. Conclusion: The present study
describes an efficient RNAi system for gene silencing that is specific to tumor
cells using the hTERT promoter. Suppression of Bcl-2 by using this system sensitized
HeLa cells to 5-fluorouracil. This system may be useful for RNAi therapy.},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/3785}
}