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

• Дата: 2008
• Автори: Minchenko, D.O., Bobarykina, A.Y., Ratushna, O.O., Marunych, R.Y., Tsuchihara, K., Moenner, M., Caro, J., Esumi, H., Minchenko, O.H.
• Формат: Стаття
• Мова: English
• Опубліковано: Інститут біохімії ім. О.В. Палладіна НАН України 2008
• Назва видання: Біотехнологія
• Теми: Експериментальні статті
• Онлайн доступ: http://dspace.nbuv.gov.ua/handle/123456789/28152
• Назва журналу: Digital Library of Periodicals of National Academy of Sciences of Ukraine
• Цитувати: 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 // Біотехнологія. — 2008. — Т. 1, № 4. — С. 49-56. — Бібліогр.: 47 назв. — англ.

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.