新加良附方抑制人胃癌细胞增殖的分子机制

Molecular Mechanisms of Xinjialiang Fufang in Inhibiting Human Gastric Cancer Cell Proliferation

目的:探讨新加良附方及其主要成分薯蓣皂苷药物血清对人胃癌细胞BGC-823的增殖抑制作用及可能的作用机制。方法:制备新加良附方及薯蓣皂苷药物血清,并作用于胃癌细胞株BGC-823,使用MTS方法检测细胞增殖抑制率;采取实时PCR方法观察新加良附方及薯蓣皂苷药物血清对人胃癌细胞BGC-823中caspase-3及p53表达的影响;采取实时PCR方法检测胃癌细胞中miR-34a表达水平。结果:新加良附方药物血清及薯蓣皂苷药物血清对胃癌细胞BGC-823有明显的增殖抑制作用,并且其作用与时间正相关。新加良附方对胃癌细胞的增殖抑制作用在48 h、72 h高于其主要成分薯蓣皂苷;人胃癌BGC-823细胞中caspase-3的表达明显低于其在正常胃黏膜上皮细胞GES-1中的水平,p53基因表达水平高于正常GES-1细胞中的p53表达水平。新加良附方药物血清及薯蓣皂苷药物血清作用于BGC-823细胞后其caspase-3表达水平均明显上升,且随着时间增加其表达量升高;p53基因表达水平下降,并且与时间负相关。新加良附方与其主要成分薯蓣皂苷相比较,两者对caspase-3、p53的调控作用无明显差异;胃癌细胞中miR-34a表达水平高于正常胃黏膜上皮细胞GES-1。结论:新加良附方及其主要成分薯蓣皂苷可以显著抑制胃癌BGC-823细胞的生长,新加良附方对胃癌细胞的抑制作用高于其主要成分薯蓣皂苷。新加良附方及薯蓣皂苷抑制胃癌细胞增殖作用可能与增加凋亡相关蛋白caspase-3表达,抑制突变型p53表达相关。miR-34a可能发挥癌基因的作用,应明确其作用并进行下一步研究。

Objective： To explore the inhibitory effect of Xinjialiang Fufang serum and Dioscin serum on the proliferation of human gastric cancer cells BGC-823 and the possible mechanism of their action. Methods： To prepare Xinjialiang Fu Fang serum and Dioscin serum and act on the human gastric cancer cells BGC-823 and detect the cell proliferation inhibitory rate using MTS assay. To observe the effect of Xinjialiang Fufang serum and Dioscin serum on the expression of caspase-3 and p53 in human gastric cancer cells BGC-823 using RT-PCR method. To detect the expression level of miR-34 a in gastric cancer cell using RT-PCR method. Results： Xinjialiang Fufang serum and Dioscin serum have significant cell proliferation inhibitory effect against the human gastric cancer cells BGC-823 and its effect is directly proportional to time. The proliferation inhibitory effect of Xin Jia Liang Fu Fang serum is better than Dioscin serum at 48 h and 72 h. The expression level of caspase-3 in human gastric cancer cells BGC-823 is lower than the normal gastric mucosa epithelial cells GES-1,while its p53 expression level is higher than the normal GES-1 cell. After the XinJia Liang Fu Fang serum and Dioscin serum have acted on the BGC-823 cell,its caspase-3 gene expression level significantly increased and the expression level has increased with time,while the p53 gene expression level decreased and is inversely proportional to time. The regulation of caspase-3 and p53 gene between Xin Jia Liang Fu Fang serum and Dioscin serum has no statistical difference. The expression level of miR-34 a in gastric cancer cell is higher than the normal gastric mucosa epithelial cells GES-1. Conclusion： Xinjialiang Fufang and Dioscin can significantly inhibit the proliferation of human gastric cancer cells BGC-823 and their effect may be related to the increase of caspase-3 expression and the inhibition of mutated p53 expression. MiR-34 a might play a role of oncogene and it is necessary to study further for the next research.