黑素瘤细胞及表皮黑素细胞中酪氨酸激酶c-abl,c-kit和PDGFRα/β的表达

Expression of Tyrosine Kinases c-abl,c-kit,PDGFRα/β in Melanoma Cells and Melanocyte

目的观察酪氨酸激酶在多株黑素瘤细胞系及原代表皮黑素细胞中的表达,探讨酪氨酸激酶在黑素瘤发生发展过程中的作用。方法分离并培养人原代表皮黑素细胞,培养浅表垂直生长期黑素瘤细胞株WM793B,转移性黑素瘤细胞株A2058,WM-266-4,Hs294T,SK-MEL-5和SK-MEL-1,用RT-PCR法分别检测7株细胞中酪氨酸激酶c-abl,c-kit和PDGFRα/βmRNA的表达;Western-blot法检测7组细胞中非受体性酪氨酸激酶c-abl蛋白水平的表达。结果与表皮黑素细胞和WM793B细胞相比,5株转移性黑素瘤细胞中的c-ablmRNA及其蛋白水平的表达偏高;而PDGFRα/βmRNA表达明显偏低、c-kitmR-NA在表皮黑素细胞,A2058,WM-266-4和Hs294T中mRNA表达较高,而在WM793B,SK-MEL-5和SK-MEL-1中偏弱。结论酪氨酸激酶c-abl对黑素瘤的发生发展可能有促进作用,而PDGFRα/βmRNA的低表达可能与黑素瘤的转移有关。

Objective To observe the expression of tyrosine kinases both in melanoma cell lines and primary cutaneous melanocyte and to determine the role of tyrosine kinases in the melanoma occurrence and development. Methods Isolate and culture primary cutaneous melanocytes from homo sapiens, culture superficial vertical growth phase melanoma cell hne WM795B, and 5 metastatic melanoma cell lines A2058, WM-2664, Hs294T, SK-MEL-5 ,SK-MEL-1. The mRNA expression of tyrosine kinase c-abl, c-kit, PDGFRα/β in pri- mary cutaneous melanocytes and melanoma cell lines were detected by RT-PCR and nonreceptor tyrosine ki- nase c-abl expression was also detected by western blot. Results C-abl mRNA and protein expression in 5 metastatic cell lines were higher than that in primary cutaneous melanocytes and superficial vertical growth phase melanoma cell line WM793B, meanwhile, PDGFRα/β mRNA levels were markedly lower in 5 meta- static melanoma cell lines. The mRNA expression of c-kit in primary cutaneous melanocytes, A2058, WM- 266-4, Hs294T were slightly higher than that in WM793B, SK-MEL-5 and SK-MEL-1. Conclusion Tyro- sine kinase c-abl could promote melanoma occurrence and development, low mRNA expression of PDGFRα/β may be related with melanoma metastasis .