原花青素对食管癌细胞增殖及糖酵解的影响及作用机制研究

摘要

Objective To investigate the effect and mechanism of proanthocyanidins on the proliferation and glycoly-sis of esophageal cancer cells.Method Esophageal cancer cells OE33,CP-C and Eca109 were treated with 0,60,120, 180,240 and 300 μmol/L proanthocyanidins respectively,and the cell survival was detected with MTT assay for calculat-ing half maximal inhibitory concentration (IC50), which was subsequently used to treat Eca109 cells, and then the ATP content,pyruvate kinase(PK)activity,hexokinase(HK)activity and lactic acid content in the supernatant were detected using testing kits,the expression levels of Akt,p-Akt,STAT3 and p-STAT3 were detected by Western blot.Result Pro-anthocyanidins can inhibit the proliferation of esophageal cancer cells OE33,CP-C and Eca109 in a concentration-depen-dent manner,the IC50was(285.48±3.58)μmol/L,(291.00±3.36)μmol/L,(237.95±4.91)μmol/L,respectively,and the last concentration demonstrated the maximum inhibitory effect on Eca109 cell line cells,and were selected as the study con-centration. After treatment with 240 μmol/L proanthocyanidins on Eca109 cell line cells, the ATP content, PK activity, HK activity and secretion of lactic acid levels were significantly reduced,and the expression levels of p-Akt and p-STAT3 also declined obviously as compared with the treatment group of 0 μmol/L(P<0.05).Conclusion Proanthocyanidins can inhibit the proliferation of esophageal cancer cells, influencing the glycolytic pathway of esophageal cancer cells, of which the mechanism may be involved with Akt and STAT3 signaling pathways.%目的 探讨原花青素对食管癌细胞株细胞增殖及糖酵解的影响及作用机制.方法 采用0、60、120、180、240、300 μmol/L的原花青素作用于食管癌细胞株OE33、CP-C、Eca109后,噻唑蓝检测细胞存活情况,计算半数抑制浓度,筛选增殖抑制作用最大的Eca109细胞继续研究.用半数抑制浓度的原花青素作用于Eca109细胞后,分别用试剂盒检测三磷酸腺苷(ATP)含量、丙酮酸激酶活性、己糖激酶活性及上清中乳酸含量,Western blot法检测蛋白激酶B(Akt)、磷酸化的Akt(p-Akt)、信号转导与转录因子3(STAT3)、磷酸化的STAT3(p-STAT3)的表达水平.结果 原花青素能够呈浓度依赖地抑制食管癌细胞株OE33、CP-C、Eca109的增殖,其半数浓度依次为(285.48±3.58)μmol/L、(291.00±3.36)μmol/L、(237.95±4.91)μmol/L,其对Eca109细胞增殖抑制作用最大,后续选用240 μmol/L的原花青素作用于Eca109细胞.240 μmol/L原花青素作用后,Eca109细胞的ATP含量、丙酮酸激酶活性、己糖激酶活性和分泌的乳酸水平均较0 μmol/L组降低(P﹤0.05),p-Akt、p-STAT3水平也较0 μmol/L组降(P﹤0.05).结论 原花青素能够抑制食管癌细胞增殖,影响食管癌细胞糖酵解途径,Akt、STAT3信号通路可能是其作用机制之一.