五氯联苯PCB126对人肝癌细胞SMMC-7721有氧糖酵解的影响

发布时间：2019-04-23 08:30

【摘要】：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(P0.05);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(P0.05)。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(P0.01)。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。
[Abstract]:Objective to investigate the effect and mechanism of pentachlorobiphenyl (PCB126) on aerobic glycolysis of human hepatoma cells. Methods SMMC-7721, cells were treated with 10 ~ (- 11), 10 ~ (- 10), 10 ~ (- 9), 10 ~ (- 8), 10 ~ (- 7) mol/L PCB126 for 24 h and 48 h, respectively. 0.1%DMSO was used as control group. Glucose content and lactic acid production in culture medium were detected by kit, pyruvate kinase M2 (PKM2) and key factors in aerobic glycolysis pathway, glucose transporter 1 (GLUT1) and lactate dehydrogenase (LDHA), were detected by real-time fluorescence quantitative PCR. Expression of pyruvate dehydrogenase kinase (PDK). The effects of PKM2, on glucose content, lactic acid production and the expression of key factors in aerobic glycolysis were detected by RNA interference technique. Results compared with the control group, SMMC-7721 cells were treated with 10 ~ (- 10), 10 ~ (- 9), 10 ~ (- 8) mol/L PCB126 for 48 h. The difference was statistically significant (P0.05); Compared with the control group, 48 h treatment had no significant effect on cell survival rate, and 72 h cell survival rate significantly increased. PCB126 exposure significantly increased the levels of PKM2,GLUT1,LDHA and PDK mRNA, the difference was statistically significant (P0.05). Compared with the 10 ~ (- 9) mol/L PCB126 group, the glucose content in the culture medium increased, the lactic acid production decreased, and the levels of GLUT1,LDHA and PDK mRNA decreased significantly after PKM2 shRNA knockout of PKM2 (P0.01). Conclusion PCB126 can up-regulate the expression of GLUT1,LDHA and PDK through PKM2 and promote aerobic glycolysis, which may be related to PCB126 promoting the development of HCC.
【作者单位】： 山西大学生物技术研究所化学生物学与分子工程教育部重点实验室;
【基金】：国家自然科学基金(21207084);国家自然科学基金(31271516) 山西省自然科学基金(2014011027-5) 高等学校科技创新项目(2016122)
【分类号】：R994.6
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本文编号：2463298