木鳖子对羟基桂皮醛对黑素瘤B16细胞分化的影响及其机制

目的 探讨木鳖子Momrodica cochinchinensis中单体化合物,尤其是对羟基桂皮醛诱导黑素瘤B16细胞分化的作用.方法 以对羟基桂皮醛、松柏醛、对羟基苯甲醛、3-甲氧基对羟基苯甲醛和ligballinol处理B16细胞48 h后,磺酰罗丹明B(SRB)法检测细胞生长抑制率.以对羟基桂皮醛处理B16细胞24、48、72 h后,倒置相差显微镜和经瑞氏-吉姆萨染色法观察细胞形态;比色法检测给药后48 h B16细胞中黑色素量和酪氨酸酶活性;RT-PCR法检测酪氨酸酶基因表达.结果 5个单体化合物对B16细胞生长均具有抑制作用,其中对羟基桂皮醛对细胞生长的抑制作用最为明显,且具有浓度和时间相关性.对羟基桂皮醛处理B16细胞48 h后,经染色观察到细胞呈树突样生长,表现为典型的细胞分化形态特征,黑色素的量显著增加,酪氨酸酶活性显著增强,上述作用与对照组相比差异显著(P＜0.05).以对羟基桂皮醛处理B16细胞6、12、24h后,酪氨酸酶、酪氨酸相关蛋白-1和酪氨酸相关蛋白-2的基因表达明显上调(P＜0.01),且呈时间相关性.结论 对羟基桂皮醛能够抑制黑素瘤B16细胞增殖,其机制与诱导B16细胞分化相关.

Effect of p-hydroxylcinnamaldehyde on differentiation of melanoma cells and its mechanism

Objective To explore the monomer compounds in the seeds of Momrodica cochinchinensis and to study the differentiation of mouse melanoma B16 cells induced by p-hydroxylcinnamaldehyde (PHC). Methods After being treated by five kinds of compounds [PHC, coniferylaldehyde, p-hydroxylbenzaldehyde (PHB), 3-O-methoxyaniline-p-hydroxylbenzaldehyde, and ligballinol] for 48 h, the inhibitory rate of B16 cell growth was measured by sulforhadamine B (SRB); Morphological changes of B16 cells induced by PHC for 24, 48, and 72 h were observed by Giemsa staining and phase contrast microscope; Melanin content and the activity of tyrosinase in B16 cells 48 h after the administration were assessed by colorimeter. The expression of tyrosinase mRNA was detected by RT-PCR. Results All the five compounds had the inhibitory effect on the B16 cells. Among them, PHC showed the strongest effect in the dose- and time-dependent manner; PHC could induce B16 cells dendritic growth 48 h after the treatment, and the morphological changes were typically differentiated; PHC also increased the melanin production and the activity of tyrosinase. There was a significant difference compared to the control group (P < 0.05). After treated by PHC for 6, 12, and 24 h, the expression levels of tyrosinase mRNA, tyrosinase 1 mRNA, and tyrosinase 2 mRNA were significantly increased (P < 0.01) in a time-dependent manner. Conclusion PHC could inhibit the proliferation of B16 cells and the mechanism is related to the differentiation of B16 cells.