苦参碱调节胃癌细胞糖代谢改变其生物学特性的机制

Mechanisms of matrine regulating glucose metabolism in gastric cancer cells and changing their biological characteristics

目的研究苦参碱(Matrine,Mat)通过调节胃癌细胞HGC-27糖代谢从而改变其转移侵袭特性的作用机制。方法体外培养HGC-27细胞,将HGC-27细胞分为对照组和低、中、高剂量实验组,对照组为常规培养的细胞,低、中、高剂量实验组用5,10,20μmol·L-1苦参碱干预。以CCK-8法检测细胞活力的改变,以流式细胞术检测细胞周期的改变,以克隆形成实验检测HGC-27的集落形成能力,以划痕实验检测细胞迁移能力,以Transwell小室检测细胞侵袭能力,以蛋白质印迹(Western blot)法检测糖酵解关键酶己糖激酶(HK)、丙酮酸脱氢酶(PDH)、柠檬酸合酶(CS)的表达,以试剂盒检测培养基中乳酸的含量。结果苦参碱干预后,HGC-27细胞活力下调,与对照组相比,差异有统计学意义(P<0.05),同时周期得到抑制,G0/G1期细胞比例上调。对照组和低、中、高剂量实验组细胞集落数分别为187.87±32.43,129.43±23.55,75.44±13.54,25.43±7.76;侵袭细胞数目分别为154.65±22.43,100.43±19.55,65.54±8.87,48.76±3.43,差异均有统计学意义(均P<0.05)。细胞中糖酵解关键酶HK、PDH和CS的表达下调,且具有剂量依赖性,与对照组相比差异均有统计学意义(均P<0.05),同时苦参碱干预后细胞中乳酸的水平下调(P<0.05)。结论苦参碱对于HGC-27细胞的转移侵袭能力具有抑制作用,苦参碱可以通过改善HGC-27细胞的糖代谢水平抑制胃癌细胞的转移和侵袭,这是苦参碱抗肿瘤作用的机制之一。

Objective To study the mechanism of matrine(Mat) in regulating the glycometabolism of gastric cancer cell line HGC-27 to alter its metastasis and invasion. Methods HGC-27 cells were cultured in vitro, and were divided into control group and low, medium and high dose experimental groups. Control group was conventionally cultured cells. Low, medium and high dose experimental groups were intervented with 5, 10, 20 μmol·L-1 Mat. CCK-8 method was used to detect the changes of cell viability, flow cytometry was used to detect the changes of cell cycle, clone formation test was used to detect the colony-forming ability of HGC-27, scratch test was used to detect the cell migration ability, transwell chamber was used to detect cell invasion ability. Western blot was used to detect the expression of hexokinase(HK), pyruvate dehydrogenase(PDH), citrate synthase(CS), the content of lactic acid in culture medium was detected by kit. Results After Mat intervention,the viability of HGC-27 cells were decreased significantly compared with control group( P < 0. 05). At the same time,the cell cycle was inhibited and the proportion of G0/G1 cells increased. The colony numbers in control group and low,medium and high dose experimental groups were 187. 87 ± 32. 43,129. 43 ± 23. 55,75. 44 ± 13. 54 and 25. 43 ± 7. 76,the number of invasive cells were 154. 65 ± 22. 43,100. 43 ± 19. 55,65. 54 ± 8. 87 and 48. 76 ± 3. 43,all with significant difference( all P < 0. 05). The expression of glycolysis key enzymes HK,PDH and CS were down-regulated in a dose-dependent manner,which was significantly different compared with control group( P < 0. 05),and the level of lactic acid was down-regulated after Mat intervention( P < 0. 05). Conclusion Matrine can inhibit the metastasis and invasion of HGC-27 cells. Mat can inhibit the metastasis and invasion of gastric cancer cells by improving the glycometabolism level of HGC-27 cells,which is one of the mechanisms of anti-tumor effect of matrine.