海藻多糖通过下调肝癌细胞Hep3B糖酵解途径抑制细胞增殖和迁移
发布时间：2019-07-26 19:24
【摘要】：目的:探讨海藻多糖(algal polysaccharides,AP)对肝癌细胞Hep3B的增殖和迁移的影响及其可能的作用机制,为治疗肝癌提供新的思路。方法:(1)比较正常细胞与癌细胞中糖酵解(embden-meyerhof-parnas,EMP)途径的表达差异,用紫外分光光度计比色法检测EMP限速酶酶活力:己糖激酶(hexokinase,HK)、丙酮酸激酶(pyruvate kinase,PK),用乳酸测定试剂盒测定EMP产物:乳酸。(2)AP处理Hep3B后,检测EMP表达水平变化。(3)AP处理细胞后,检测肝癌细胞Hep3B的增殖和迁移能力及对上皮细胞-间充质转化(EMT)的影响,方法包括MTT、q PCR和Transwell小室实验。(4)MTT、Transwell和q PCR检测HK抑制剂3-溴丙酮酸(3-bromopyruvate,3-BrPA)对肝癌Hep3B细胞的活力和迁移能力及EMT的影响。(5)Western blot和相关试剂盒检测AP与3-BrPA分别处理细胞时,对EMP水平及Akt通路的影响。(6)AP联合3-BrPA处理Hep B3后,检测HK和Akt信号通路表达水平及细胞活力和迁移的变化。结果:(1)癌细胞(Hep3B、He La、SW480)EMP代谢水平均高于正常肝细胞(HL02),其中Hep3B差异最为显著。(2)AP能抑制Hep B细胞EMP代谢水平,且抑制程度随着AP浓度升高而升高,存在浓度依赖性。(3)AP可抑制肝癌细胞Hep3B的增殖和迁移,且抑制EMT的发生。AP浓度为200mg/ml,处理时间为48h时,生长抑制率达(55±2.8)%,细胞迁移数为对照组的(32±2.9)%;(4)3-BrPA可下调Hep3B细胞HK活性,并抑制细胞活力与迁移,且抑制EMT的发生。200μmol/L 3-BrPA作用细胞48h后,与对照组相比,细胞活性下降了(52±5.8)%(P0.001),细胞迁移数下降了(48±6.1)%(P0.01)。(5)AP和3-BrPA均下调EMP代谢水平,且抑制Akt信号通路。(6)AP与3-BrPA联合处理Hep3B后,HK表达下降,显著抑制Akt信号通路,细胞活性和迁移能力均较AP单用组显著下降(P0.05)。结论:肝癌细胞Hep3B EMP代谢水平高于正常肝细胞,AP可下调Hep3B细胞EMP代谢水平,低EMP代谢水平可能通过下调EMT和Akt信号通路抑制Hep3B的增殖和迁移。当AP和3-BrPA联合应用时,抑制肝癌迁移和增殖效果更明显。
[Abstract]:Objective: to investigate the effect of seaweed polysaccharide (algal polysaccharides,AP) on the proliferation and migration of HCC cell line Hep3B and its possible mechanism, so as to provide a new idea for the treatment of HCC. Methods: (1) the expression difference of glycolysis (embden-meyerhof-parnas,EMP) pathway between normal cells and cancer cells was compared. The activities of EMP rate-limiting enzyme were detected by ultraviolet spectrophotometer: hexokinase (hexokinase,HK), pyruvate kinase (pyruvate kinase,PK), EMP product: lactic acid was measured by lactic acid assay kit. (2) after Hep3B treatment with AP, the expression level of EMP was detected. (3) after AP treatment, the expression level of EMP was detected. The proliferation and migration ability of hepatocellular carcinoma cell line Hep3B and the effect of MTT,Transwell and Q PCR on the proliferation and migration of HCC cells and the transformation of EMT into epithelial cells were detected by MTT,Transwell and Q PCR. (5) Western blot and related kits were used to detect the effect of AP and 3-BrPA on the activity and migration of HCC Hep3B cells, and the effect of HK inhibitor 3 bromopyruvate, 3 鈮，
本文编号：2519751
[Abstract]:Objective: to investigate the effect of seaweed polysaccharide (algal polysaccharides,AP) on the proliferation and migration of HCC cell line Hep3B and its possible mechanism, so as to provide a new idea for the treatment of HCC. Methods: (1) the expression difference of glycolysis (embden-meyerhof-parnas,EMP) pathway between normal cells and cancer cells was compared. The activities of EMP rate-limiting enzyme were detected by ultraviolet spectrophotometer: hexokinase (hexokinase,HK), pyruvate kinase (pyruvate kinase,PK), EMP product: lactic acid was measured by lactic acid assay kit. (2) after Hep3B treatment with AP, the expression level of EMP was detected. (3) after AP treatment, the expression level of EMP was detected. The proliferation and migration ability of hepatocellular carcinoma cell line Hep3B and the effect of MTT,Transwell and Q PCR on the proliferation and migration of HCC cells and the transformation of EMT into epithelial cells were detected by MTT,Transwell and Q PCR. (5) Western blot and related kits were used to detect the effect of AP and 3-BrPA on the activity and migration of HCC Hep3B cells, and the effect of HK inhibitor 3 bromopyruvate, 3 鈮，
本文编号：2519751
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