The Expression Of Hypoxia-Inducible Factor-1a And PK-M2in Cholangiocarcinoma Cells And Its Effect In Glycolysis

Objective:To examine the expression change of HIF-1a and PK-M2in protein levels in QBC939cells under chemical hypoxia-induced by CoCl2in vitro, meanwhile to explore the effects and mechanism of YC-1about the Inhibition effect of HIF-1aand tumor glycolysis.Methods:QBC939cells were divided as control group and experiment group. The former were cultured with normal condition, the later were treated by YC-1(150μmol/L). Western blot technique were used to examine the change of HIF-1a and PK-M2protein levels in QBC939cells in different concentrations (0,100,150,200,250μmol/L) of CoCl2and different periods (6h,12h,24h,48h) in hypoxia and pre-post of YC-1. The activity of lactic acid(LA) concentration in the culture supernatant was tested with spectrophotometer method in the24h group.Results:①Under chemical hypoxia-induced by different CoCl2in vitro, the expression of HIF-1a and PK-M2in the QBC939cells gradually increased as the concentration of CoCl2increased and hypoxia time extended (P<0.05), and the expression of HIF-1a reached a peak at the group of150μmol/L CoCl2and the group of12h.②Under chemical hypoxia-induced by different levels of CoCl2(0、100、150、200、250μmol/L) in the24h groups in vitro, the lactic acid secretion of QBC939cells gradually increased as the concentration of CoCl2increased, compared with that under normal oxygen condition, the lactic acid secretion of QBC939cells obviously increased (P<0.05).③Cultivated QBC939cells in the condition of150μmol/L YC-1and different levels of CoCl2, both the expression of HIF-1a and PK-M2decreased to some extent. The expression of HIF-1a obviously decreased in the groups of150,250μmol/L CoCl2and the groups of12,24,48h, and the difference was statistic significant (P<0.05). The expression of PK-M2obviously decreased in the groups of100,150,200pmol/L CoCl2and the groups of6,24,48h, and the difference was statistic significant (P<0.05).④Cultivated QBC939cells24h in the condition of150μmol/L YC-1and different levels of CoCl2, the lactic acid secretion obviously decreased in the groups of100,150,200,250/anol/L CoCl2, and the difference was statistic significant (P<0.05).Conclusions:①QBC939cell line of hypoxia, there is up-regulation HIF-laand PK-M2, between HIF-la and PK-M2are positively correlated relationship, they are both enhance the effect of tumor glycolysis.②In QBC939cell line, YC-1can effectively inhibit the activity of HIF-1a, further can inhibit the expression of PK-M2, and then effectively reduce the effect level of glycolysis.