The impact of 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase-3 gene on the proliferation of prostate cancer cells

Abstract

Objective To investigate the expression of 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase-3 (PFKFB3) gene in the prostate cancer tissues and its impact on the proliferation of prostate cancer cells PC3 and DU145. Methods Immunohistochemical staining was used to detect the expression of PFKFB3 in the prostate cancer tissues and the adjacent tissues (antibody concentration 1∶50). Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR, 40 cycles, 188 bp) and Western blotting [30 μl, 10% sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE)] were applied to detect the mRNA and protein expression levels of PFKFB3 expression in the prostate cancer cells and prostate epithelial cells. After knocking out PFKFB3 gene, plate cloning (40 μmol/L H2O2) formation assay and cell growth assay [20 μl methyl thiazol tetrazolium (MTT)] were applied to measure the cell proliferation. Results The expression of PFKFB3 in the prostate cancer tissues (2.42±0.16) was significantly higher than that in the adjacent tissues (1.38±0.10, P=0.031). The mRNA and protein expression levels of PFKFB3 in prostate cancer cells were significantly higher than those in the prostate epithelial cells (P=0.015), 2.0-2.6 times and 2.5-4.5 times respectively. When knocking out the PFKFB3 gene, cloning formation assay showed that the cloning formation rate decreased as 22%-35% compared to the control (P=0.018). Cell growth assay showed that the proliferation of prostate cancer cells was inhibited as 28%-30% at day 4 (P=0.026). Conclusion PFKFB3 gene is highly expressed in prostate cancer cells and prostate cancer tissues. PFKFB3 plays a promotional role in the prostate cancer cells. Key words: 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase-3; Prostate cancer; Proliferation