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目的探讨Wnt5a-Ca2+-CN-NFAT通路主要信号分子在正常人黑素细胞及不同黑素瘤细胞系中的激活状态。方法选取正常人原代黑素细胞(MC)及来源于原位(WM793B)、侵袭性(LIBR)及转移性(SK-MEL-5)黑素瘤细胞系和色素痣、黑素瘤组织,采用逆转录-聚合酶链反应(RT-PCR)、蛋白质印迹法(Western blot)、激光共聚焦及免疫组化法检测细胞及组织中该通路主要信号分子的表达情况。结果与正常人黑素细胞相比,在不同黑素瘤细胞系中该通路主要信号分子Wnt5a、散乱蛋白2(DVL2)、钙调神经磷酸酶(calcineurin,CN)、活化T细胞核因子2(NFAT2)、环氧合酶-2(COX-2)、血管内皮细胞生长因子A(VEGF-A)、黏着斑蛋白(vinculin)及波形蛋白(vimentin)明显高表达,且Ca2+浓度较高;CN-B、NFAT2在色素痣中均为阴性表达,而在黑素瘤组织中多数呈阳性表达(P<0.001)。在侵袭性及转移性黑素瘤中的表达强度明显高于原位黑素瘤。结论在黑素瘤的发生、发展过程中,Wnt5aCa2+-CN-NFAT信号通路可能被激活,并在黑素瘤的侵袭和转移中发挥重要作用。
Objective To investigate the activation of major signaling molecules of Wnt5a-Ca2 + -CN-NFAT pathway in normal human melanocytes and different melanoma cell lines. Methods Normal human primary melanocytes (MCs) and melanoma cell lines derived from in situ (WM793B), invasive (LIBR) and metastatic (SK-MEL-5) Reverse transcription-polymerase chain reaction (RT-PCR), Western blot, laser scanning confocal microscopy and immunohistochemistry were used to detect the expression of major signaling molecules in the cells and tissues. Results Compared with normal melanocytes, Wnt5a, caspase-2, calcineurin (CN) and NFAT2 in different melanoma cell lines ), COX-2, VEGF-A, vinculin and vimentin were significantly higher than those in control group, and the concentration of Ca2 + B, NFAT2 were negative in pigmented nevus, while in melanoma tissues were mostly positive (P <0.001). The expression intensity in invasive and metastatic melanoma was significantly higher than that in situ melanoma. Conclusion The Wnt5aCa2 + -CN-NFAT signaling pathway may be activated during the development of melanoma and play an important role in the invasion and metastasis of melanoma.