DOMINANT-NEGATIVE CONSTRUCTS OF HUMAN 6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE-2,6-BISPHOSPHATASE-3 AND 4: EFFECT ON THE EXPRESSION OF ENDOGENOUS 6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE-2,6-BISPHOSPHATASE mRNA D. O. Minchenko, A. Y. Bobarykina, O. O. Ratushna, R. Y. M

Details: Category: 4_2008(en); Hits: 163

ISSN 1995-5537

"Biotechnology" journal Vol. 1, No. 4, 2008
Р. 49-56, Bibliography 47, English.
Universal Decimal classification 577.112:616

DOMINANT-NEGATIVE CONSTRUCTS OF HUMAN
6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE-2,6-BISPHOSPHATASE-3 AND 4: EFFECT ON THE EXPRESSION OF ENDOGENOUS 6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE-2,6-BISPHOSPHATASE mRNA

D. O. Minchenko, A. Y. Bobarykina, O. O. Ratushna, R. Y. Marunych, K. Tsuchihara,
M. Moenner, J. Caro, H. Esumi, O. H. Minchenko

Palladin Institute of Biochemistry of the National Academy of Science of Ukraine, Kyiv

Research Center for Innovative Oncology of the National Cancer Center Hospital East,Kashiwa, Japan

INSERM U920 Molecular Mechanisms of Angiogenesis Laboratory,

University Bordeaux 1, Talence, France

Department of Medicine,

Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA, USA

Expression of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB), a key regulatory enzyme of glycolysis, is significantly increased in different malignant tumors provides a potential mechanism of enhanced glycolysis and cancer cell proliferation.

We created dominant-negative constructs of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 and -4 (dnPFKFB-3 and dnPFKFB-4) cDNA for suppression of strongly enhanced expression endogenous PFKFB-3 and PFKFB-4. We introduce point mutation in ATP-binding domain of 6-phosphofructo-2-kinase part of PFKFB-3 as well as PFKFB-4 cDNA for suppression of 6-phosphofructo-2-kinase activity in the products of dnPFKFB-3 and dnPFKFB-4 expression.

Cancer cells were stable transfected with these dominant-negative constructs for suppression of endogenous PFKFB-3 and PFKFB-4 expression and cell proliferation. We have shown that PFKFB-3 expression in pancreatic cancer cell line Panc1, stable transfected by dnPFKFB-3, was significantly reduced in normal as well as in hypoxic conditions. Pancreatic cancer cells proliferation, stable transfected by dnPFKFB-3 or dnPFKFB-4, was also reduced. Results of this investigation demonstrate possibility to apply the dominant-negative constructs of PFKFB-3 and PFKFB-4 for suppression of glycolysis and tumor cells proliferation via reduction of endogenous PFKFB expression.

Key words: PFKFB-4 mRNA, PFKFB-3 mRNA, dominantnegative constructs, cancer cells.