PMID- 17328665
OWN - NLM
STAT- MEDLINE
DCOM- 20070405
LR  - 20181201
IS  - 1044-5498 (Print)
IS  - 1044-5498 (Linking)
VI  - 26
IP  - 2
DP  - 2007 Feb
TI  - Combined p14ARF and antisense EGFR potentiate the efficacy of adenovirus-mediated 
      gene therapy in laryngeal squamous cell carcinoma (LSCC).
PG  - 71-9
AB  - The tumor suppressor p14(ARF) and protooncogene epidermal growth factor receptor 
      (EGFR) play an important role in the development of laryngeal squamous cell 
      carcinoma (LSCC). We explored the inhibition of proliferation and induction of 
      differentiation in human larynx cancer cells (Hep-2) in vitro when p14(ARF) 
      couples with antisense complementary DNA of EGFR to transfect into Hep-2 cells 
      via the AdEasy-1 vector system. In vitro studies, using standard isobologram 
      analyses, identified whether Ad-antisense EGFR is synergistic with Ad-14(ARF). To 
      evaluate the cytotoxicity of these agents the gold standard clonogenic survival 
      assay was used. Western blotting analyses, 
      3'(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and 
      flow cytometer (FCM) analysis was used to detect protein expression, 
      proliferation, and cell cycle distribution of Hep-2 cells, respectively. 
      Meanwhile, empty vector and PBS were set as a control. The activity of 
      proliferation of Hep-2 cells was inhibited markedly by infection of Ad-p14(ARF) 
      combined with Ad-antisense EGFR compared with Ad-p14(ARF) or Ad-antisense EGFR 
      alone (P = 0.001, P = 0.002, respectively), with Ad-sense EGFR (P = 0.0005), with 
      vector control (Ad-Ctrl) (P = 0.0001), and with PBS (P = 0.0001). FCM revealed 
      that the proportion in the G(0)/G(1) phases increased by up to 86.9% when 
      Ad-p14(ARF) was associated with Ad-antisense EGFR to transfect Hep-2 cells. A 
      weakened expression of EGFR protein and P14 (ARF) protein overexpression was 
      observed. Our study in vitro indicated that association of Ad-p14(ARF) with 
      Ad-antisense EGFR remarkably inhibited activity of proliferation and inducted 
      differentiation of Hep-2 cells. Therefore, not only EGFR, but also p14(ARF), 
      plays a major role in the genesis and in modulating cell growth and 
      differentiation of LSCC, and their synergistic effect was obvious. An effective 
      potential target of gene therapy to prevent LSCC proliferation was provided.
FAU - Xian, Junming
AU  - Xian J
AD  - Department of Otolaryngology, West China Hospital, Sichuan University, Chengdu, 
      People's Republic of China.
FAU - Lin, Yinghe
AU  - Lin Y
FAU - Liu, Yafeng
AU  - Liu Y
FAU - Gong, Ping
AU  - Gong P
FAU - Liu, Shixi
AU  - Liu S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - DNA Cell Biol
JT  - DNA and cell biology
JID - 9004522
RN  - 0 (DNA, Antisense)
RN  - 0 (Tumor Suppressor Protein p14ARF)
RN  - EC 2.7.10.1 (ErbB Receptors)
SB  - IM
MH  - Adenoviridae/*genetics
MH  - Carcinoma, Squamous Cell/*pathology/therapy
MH  - Cell Cycle
MH  - Cell Differentiation
MH  - Cell Line, Tumor
MH  - Cell Proliferation
MH  - Cell Survival
MH  - DNA, Antisense/*genetics
MH  - ErbB Receptors/biosynthesis/*genetics
MH  - Genetic Therapy
MH  - Genetic Vectors
MH  - Humans
MH  - Laryngeal Neoplasms/*pathology/therapy
MH  - *Transfection
MH  - Tumor Suppressor Protein p14ARF/biosynthesis/*genetics
EDAT- 2007/03/03 09:00
MHDA- 2007/04/06 09:00
CRDT- 2007/03/03 09:00
PHST- 2007/03/03 09:00 [pubmed]
PHST- 2007/04/06 09:00 [medline]
PHST- 2007/03/03 09:00 [entrez]
AID - 10.1089/dna.2006.0533 [doi]
PST - ppublish
SO  - DNA Cell Biol. 2007 Feb;26(2):71-9. doi: 10.1089/dna.2006.0533.