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目的探讨五氯联苯PCB126对人肝癌细胞有氧糖酵解的影响和机制。方法取人肝癌细胞SMMC-7721,用浓度分别为10~(-11)、10~(-10)、10~(-9)、10~(-8)、10~(-7) mol/L PCB126分别处理24、48 h,并以0.1%DMSO作为对照组。用试剂盒检测培养基中葡萄糖含量和乳酸生成量,以实时荧光定量PCR法检测丙酮酸激酶M2(PKM2)和有氧糖酵解通路中关键因子葡萄糖转运蛋白1(GLUT1)、乳酸脱氢酶(LDHA)、丙酮酸脱氢酶激酶(PDK)的表达。用RNA干扰技术敲减PKM2,检测其对培养基中葡萄糖含量、乳酸生成量和有氧糖酵解关键因子表达的影响。结果与对照组相比,人肝癌细胞SMMC-7721经10~(-10)、10~(-9)、10~(-8)mol/L PCB126处理48 h后,培养基中葡萄糖含量明显下降,乳酸生成量明显增加,差异有统计学意义(P<0.05);与对照组相比,处理48 h后对细胞存活率无明显影响,72 h时细胞存活率明显升高。PCB126暴露可明显升高PKM2、GLUT1、LDHA和PDK mRNA水平,差异有统计学意义(P<0.05)。用PKM2 shRNA敲减PKM2后,与10~(-9) mol/L PCB126处理组相比,培养基中葡萄糖含量升高,乳酸生成量下降,GLUT1、LDHA和PDK mRNA水平明显降低,差异均有统计学意义(P<0.01)。结论 PCB126可通过PKM2上调肝癌细胞GLUT1、LDHA和PDK的表达,从而促进有氧糖酵解,这可能与PCB126促进肝癌的发展有关。
Objective To investigate the effect and mechanism of PC12 PCB126 on aerobic glycolysis of human hepatoma cells. Methods Human hepatocellular carcinoma cell line SMMC-7721 was harvested from human hepatocellular carcinoma cell line SMMC-7721 with concentrations of 10 ~ (-11), 10 ~ (-10), 10 ~ (-9), 10 ~ (-8) PCB126 were treated for 24,48 h, respectively, and 0.1% DMSO as a control group. The contents of glucose and lactate in culture medium were detected by kit, and the expressions of key enzymes such as glucose transporter 1 (GLUT1), pyruvate kinase M2 (PKM2) and lactate dehydrogenase (LDH) in aerobic glycolysis pathway were detected by real-time fluorescence quantitative PCR (LDHA), pyruvate dehydrogenase kinase (PDK) expression. PKM2 was knocked down by RNA interference and its effect on glucose content, lactate production and expression of key factors of aerobic glycolysis in culture medium was tested. Results Compared with the control group, the glucose content in the medium of SMMC-7721 cells treated with 10 ~ (-10), 10 ~ (-9), 10 ~ (-8) mol / L PCB126 for 48 h decreased significantly (P <0.05). Compared with the control group, the cell viability was not significantly affected after 48 h of treatment, and the cell survival rate at 72 h was significantly increased. PCB126 exposure significantly increased PKM2, GLUT1, LDHA and PDK mRNA levels, the difference was statistically significant (P <0.05). Compared with 10 -9 mol / L PCB126 treatment group, knockdown of PKM2 by PKM2 shRNA resulted in an increase in glucose content, a decrease in lactate production, and a significant decrease in GLUT1, LDHA and PDK mRNA levels in the medium with 10 -9 mol / L PCB126 treatment Statistical significance (P <0.01). Conclusion PCB126 can promote aerobic glycolysis by up-regulating the expression of GLUT1, LDHA and PDK in hepatoma cells via PKM2, which may be related to the promotion of hepatocarcinogenesis by PCB126.