去甲斑蝥素对抑制人A375黑素瘤细胞增殖、黑色素合成及酪氨酸酶活性的研究

Inhibition effects of norcantharidin on the proliferation, melanin synthesis and tyrosinase activity in human melanoma cell strain A375

目的以人A375黑素瘤细胞为实验模型,研究去甲斑蝥素对黑素瘤细胞的增殖、黑色素含量及酪氨酸酶活性的影响。方法培养人A375黑素瘤细胞,采用10、20、40、80μmol/L的去甲斑蝥素预处理,培养24 h和48 h后,四甲基偶氮唑蓝比色法测定细胞增殖活性;Na OH裂解法测定细胞黑色素含量;加入左旋多巴液法测定细胞酪氨酸酶活性。结果 10μmo/L去甲斑蝥素作用在人A375黑素瘤细胞24 h后,去甲斑蝥素明显的抑制该细胞的增殖活性(P<0.05)。但是对该细胞黑色素含量以及酪氨酸活性没有明显的作用(P>0.05)。20~80μmol/L的去甲斑蝥素作用在人A375黑素瘤细胞24 h后,明显抑制该细胞增殖活性、黑色素合成以及酪氨酸酶活性(P<0.05或<0.01)。20~40μmo/L的去甲斑蝥素处理人A375黑素瘤细胞24 h和48 h后,该细胞的增殖活性、黑色素含量以及酪氨酸酶活性出现不同程度降低,差异有统计学意义(P<0.05或<0.01)。结论低浓度(10μmol/L)去甲斑蝥素仅有抑制人A375黑素瘤细胞增殖的作用,而高浓度(20~80μmol/L)的去甲斑蝥素不仅具有抑制人A375黑素瘤细胞增殖的作用,还具有抑制人A375黑素瘤细胞的黑色素合成以及酪氨酸酶活性的作用。

Objective To investigate the inhibition effects of norcantharidin on the proliferation,melanin synthesis and tyrosinase activity in human melanoma cell strain A375 in vitro by using human melanoma cell strain A375 as experiment models. Methods The human melanoma cell strain A375 were cultured in vitro to establish a research model. The cells were pre-treated by norcantharidin with 10,20,40 and 80μmo/L,for 24 h and 48 h. Then the cell proliferation activity was determined by means of MTT method,the melanin content was measured by Na OH cleavage method,and the tyrosinase activity was detected by oxidative levodopa reaction. Results After the human melanoma cell strain A375 were cultured by norcantharidin（ 10μmo/L） for 24 h,the proliferation activity of cells was significantly inhibited（ P 〈 0. 05）. However,no significant effects on the content of melanin and activity of tyrosine were observed（ P 〉 0. 05）. Otherwise,after the cells were cultured by norcantharidin（ 20 ~ 80μmo/L） for 24 h,the proliferation,melanin synthesis and tyrosinase activity of human melanoma cell strain A375 were significantly inhibited（ P 〈 0. 05 or P 〈 0. 01）. Moreover,after the cells were cultured by norcantharidin（ 20 ~ 40μmo/L） for 24 h and 80 h,the proliferation activity melanin synthesis and tyrosinase activity of cells were significantly decrased at different extents in a dose-dependent manner（ P 〈 0. 05 or P 〈 0. 01）. Conclusion The low concentration of norcantharidin（ 10μmo/L） only has the inhibitary effects on the proliferation of human melanoma cell strain A375,however,the high concentration of norcantharidin（ 20 ~ 80μmo/L） can not only inhibit the proliferation of human melanoma cell strain A375,but also inhibit melanin synthesis and tyrosinase activity of human melanoma cell strain A375 in a dose-dependent manner.