活血化瘀中药对人黑素瘤细胞酪氨酸酶翻译后加工的影响

Effects of Blood Activating Chinese Herbs on Processing of Tyrosinase in Human Melanoma Cells

目的探讨活血化瘀中药对酪氨酸酶蛋白加工成熟的影响。方法体外传代培养人无色素黑素瘤细胞YUGEN8,分别加入红花、桃仁及赤芍3味中药提取液,利用蛋白免疫印迹、基因反转录扩增及内切糖苷酶酶切等方法,观察中药处理组细胞内酪氨酸酶的表达、成熟过程与阴性对照组的差异。结果与阴性对照组比,桃仁处理组细胞酪氨酸酶基因表达并未增加,酪氨酸酶蛋白80ku左右大分子量成熟片断表达增加,能够耐受内切糖苷酶酶切。结论桃仁提取物对人黑素瘤细胞内酪氨酸酶的加工成熟及稳定具有一定促进作用。

Objective To explore the contribution of traditional Chinese medicine with blood activating effects on the processing of tyrosinase （TYR）. Methods Human amelanotic melanoma cells YUGEN8 were cultured in vitro and treated with three traditional Chinese medicines including flos carthami, semen persicae and paeoniae radix. Gene and protein of TYR were analyzed by RT-PCR, Western Blot and hydrolyze with endoglycosidase H （Endo H）. The expression and maturity of TYR were observed and compared between treated cells and negative control. Results Accumulation of a 80 ku Endo Hresistant tyrosinase doublet, which represented mature glycoform of tyrosinase, was detected in melanocytes treated with Semen Persicae, although the expression of mRNA was normal, as observed by the comparison with negative control. Conclusion Semen Persicae extracts could induce tyrosinase maturation and stability in human melanoma cells.