ANGPTL4基因调控黑色素瘤细胞醛缩酶A表达水平的机制

Angiopoietin-like 4 modulates aldolase A expression in human melanoma cell in a PKC dependent manner

目的:探讨黑色素瘤细胞血管生成素样蛋白4(angiopoietin-like 4,ANGPTL4)对醛缩酶A(aldolase A,ALDOA)表达水平的影响及其机制。方法:分别扩增和沉默黑色素瘤细胞系WM-115及WM-266-4 ANGPTL4基因,运用PKC激动剂及阻断剂处理转染细胞。检测ALDOA在人黑色素瘤细胞中m RNA及蛋白质水平的表达,荧光素酶测定ALDOA基因启动子活性。结果:ANGPTL4基因扩增的黑色素瘤细胞组,ALDOA在m RNA及蛋白质的水平的表达均增高,ALDOA基因的启动子活性增加,且可被PKC阻断剂抑制(P<0.05)。而在ANGPTL4基因沉默细胞组中ALDOA在m RNA及蛋白质水平的表达均减低,ALDOA基因的启动子活性降低,且可被PKC激动剂恢复(P<0.05)。结论:ANGPTL4基因可通过PKC依赖途径从ALDOA基因转录及翻译水平上调人黑色素瘤细胞中ALDOA的表达。

Objective: To investigate the association between angiopoietin-like 4(ANGPTL4) and aldolase A(ALDOA) in human melanoma cell. Methods: Overexpression or knockdown ofANGPTL4 was performed in WM-115 or WM-266-4 cells, respectively. The expression ofANGPTL4 and ALDOA was measured by RT-PCR and Western blot, respectively. The promoter activity ofALDOA gene was determined by luciferase assay.Results: The promoter activity ofALDOA gene and the expression ofALDOA(m RNA and protein) were increased or decreased in the melanoma cells with overexpression or knockdown ofANGPTL4, which was blocked by selective protein kinase C(PKC) inhibitor or restored by PKC agonist, respectively.Conclusion: ANGPTL4 promotes ALDOA expression in human melanoma cell in a PKC dependent manner.