马钱苷抑制人恶性黑色素瘤细胞的作用及机制研究

Study on the inhibitory effect and molecular mechanism of loganin on human malignant melanoma cells

目的探索马钱苷抑制人恶性黑色素瘤细胞的作用和机制。方法将实验分为空白对照组、不同干预浓度(10、20、40、60、80 ng/m L)的马钱苷组,分别作用24、48、72 h,采用MTT方法检测细胞活性;依据药物作用48 h时的IC50浓度[(74.96±7.92)ng/m L],选取马钱苷20、40、60 ng/m L分别干预48 h,并采用RT-PCR法检测细胞内Bcl-2、Bax和Caspase-3 m RNA表达水平的变化;用Western blot检测细胞内Bcl-2、Bax和Caspase-3蛋白水平的变化。结果马钱苷具有抑制A375细胞增殖的作用,并呈时间和剂量依赖性。与空白对照组比较,马钱苷组细胞抑制凋亡因子Bcl-2的m RNA和蛋白质表达水平均明显下调(P<0.05或P<0.01),促凋亡因子Bax和Caspase-3的m RNA及蛋白质表达均明显上调(P<0.05或P<0.01)。结论马钱苷能抑制A375细胞增殖,其作用机制可能与下调细胞Bcl-2 m RNA与蛋白质的表达,上调Bax、Caspase-3 m RNA和蛋白质的表达,进而诱导细胞凋亡有关。

Objective To investigate the inhibitory effect and molecular mechanism of loganin on human malignant melanoma cells in vitro. Methods The experiment was divided into blank control group and different concentrations （10, 20, 40, 60, 80 ng/mL） of loganin groups, dealt for 24, 48, 72 h, the cells activity was detected by MTT method; according to the half-inhibitory concentration （ICs0） for 48 h [（74.96±7.92） ng/mL], 20, 40, 60 ng/mL of loganin were selected for 48 h intervention. The mRNA expressions of Bcl-2, Bax and Caspase-3 were detected by RT-PCR; the protein expressions of Bcl-2, Bax and Caspase-3 were detected by Western blot. Results Loganin has a most effectively inhibitory effect on A375 cells proliferation in a time and dose dependent manner. Compared with blank control group, the mRNA and protein expression levels of apoptosis inhibiting factor Bcl-2 in the loganin group were down-regulated significantly （P 〈 0.05 or P 〈 0.01）, and the mRNA and protein expression levels of pro-apoptotic factors Bax and Cas- pase-3 were all up-regulated, respectively （P 〈 0.05 or P 〈 0.01）. Conclusion Loganin can significantly inhibit the proliferation of A375 cells, which may be associated with inhibiting the mRNA and protein expressions of Bcl-2, im： proving the mRNA and protein expressions of Bax, Caspase-3, thus inducting apoptosis.