Expression Of Phosphofructokinase1in Nasopharyngeal Carcinoma And Influence Of The Inhibition Of PFK1by RNA Interference To The Biological Characteristic Of The Nasopharyngeal Carcinoma Cell Lines CNE2

Part I Expression of phosphofructokinase1and it's enzyme activity in nasopharyngeal carcinomaObjective:The purpose of this research is to investigate the expression of phosphofructokinase1and it's enzyme activity in nasopharyngeal carcinoma biopsy specimen, that is the key enzyme of the glycolytic pathway.Methods:61biopsy specimen were detected, including41tissues specimen from patients with nasopharyngeal carcinoma and20tissues specimen from patients with chronic nasopharyngitis as control group. Phosphofructokinase1mRNA expression was detected by RT-PCR, phosphofructokinase1protein was detected by Western-blot and it's enzyme activity was detected.Results:?It was observed that the expression levels of phosphofructokinase1mRNA, phosphofructokinase1protein and it's enzyme activity in nasopharyngeal carcinoma tissues were significantly higher compared to that in the chronic inflammatory tissues(P?0.01).?The expression levels of phosphofructokinase1mRNA, phosphofructokinase1protein and it's enzyme activity in patients with lymph node metastasis were significantly higher compared to that without lymph node metastasis(P?0.01).Conclusion:Phosphofructokinase1may be one of the molecular markers in occurrence and metastasis of nasopharyngeal carcinoma. The treatment of nasopharyngeal carcinoma with the phosphofructokinase1shRNA might become a new strategy in the future. Part II Influence of the inhibition of PFK1expression by RNA interference to the biological characteristic of the nasopharyngeal carcinoma cell lines CNE2Objective:Most cancer cells exhibit increased glycolysis, phosphofructokinase1(PFK1),which is essential for the glycolytic pathway. This research constructed a short hairpin RNA(shRNA) targeting PFK1, then analyze its effects on bionomics of CNE2.Methods::The four shRNAs targeting phosphofructokinase1(PFK1) were transfected into nasopharyngeal carcinoma cell lines CNE2. The expression of PFK1mRNA after transfected by the four shRNAs in nasopharyngeal carcinoma cell lines CNE2was detected by real time quantitative PCR(RT-PCR). After tranfected by the shRNA-PFK1with the highest inhibited rate, the expression of PFK1mRNA was detected by RT-PCR, PFK1protein was detected by Western-blot and it's enzyme activity was detected. Cell growth was observed by MTT assay, and apoptosis was detected by flow cytometry. Cell migration capacity was assessed by wound-healing assay, and cell invasion potential was evaluated by Transwell invasion assay.Results:The2-^^CT of sh-H-PFK1-507was less than the other three shRNA-PFK1, blank control group and plasmid control group, the sh-H-PFK1-507can inhibit the expression of the PFK1most obviously. After transfected by sh-H-PFKl-507, the expression levels of phosphofructokinase1mRNA, phosphofructokinase1protein and it's enzyme activity of CNE2cell in experimental group were significantly higher compared to which in the plasmid group and blank group (P?0.05). The growth of the CNE2cell in experimental group was inhibited after transfected by sh-H-PFKl-507, the absorbance value of the CNE2cell in experimental group on24h,36h,48h,72h discreased significantly compared to the cell in blank control group and plasmid control group(P?0.05). The apoptosis rate of the CNE2cell in experimental group increased obviously compared to the cell in blank control group and plasmid control group(P?0.01). The cell migration capacity and cell invasion potential of CNE2cell in experimental group were decreased significantly compared to the cell in blank control group and plasmid control group(P?0.01). Conclusion:shRNA-PFKl can reduce the expression of PFK1, inhibit the growth, migration capacity and invasion potential, and introduce the apoptosis of nasopharyngeal carcinoma cell lines CNE2. The therapy of nasopharyngeal carcinoma with the shRNA-PFK1could be a new strategy in the future.