miR-503靶向调控CXCL9抑制黑色素瘤细胞的迁移及侵袭

The miR-503 suppresses migration and invasion of melanoma cells by targeted-regulating CXCL9

摘要:

目的： 研究miR-503对人皮肤黑色素瘤细胞A375迁移、侵袭能力的影响及相关机制。方法： 实时荧光定量PCR（real-time quantitative PCR，qRT-PCR）检测黑色素瘤组织及细胞系中miR-503的表达水平；将miR-503模拟物（miR-503 mimics）或miR-503抑制物（miR-503 inhibitors）瞬时转染A375细胞，Transwell实验检测转染后细胞迁移及侵袭能力的变化；生物信息学预测miR-503与CXCL9的结合位点，双荧光素酶报告基因实验检测其靶向结合关系；将CXCL9单独转染或与miR-503共转染A375细胞后，检测细胞迁移及侵袭能力的变化。结果： 黑色素瘤组织中miR-503的表达明显低于癌旁组织（t=6.602，P=0.000），且黑色素瘤细胞（A375、SK-MEL-1及SK-MEL-5）中miR-503的表达也均明显低于人表皮黑色素细胞HEM（F=19.445，P=0.000）；A375细胞转染miR-503 mimics后迁移及侵袭能力明显受到抑制（P=0.017，P=0.049），而转染miR-503 inhibitors后迁移及侵袭能力明显增强（P=0.004，P=0.000）；CXCL9是miR-503的直接靶基因；CXCL9能促进A375细胞的迁移和侵袭（P=0.000，P=0.009），但其作用效果能被miR-503逆转。结论： miR-503通过靶向调控CXCL9而抑制黑色素瘤细胞的迁移及侵袭。

Abstract:

Objective: To investigate the effect of miR-503 on migration and invasion in human melanoma cells A375 and its mechanism. Methods: The real-time quantitative PCR (qRT-PCR) was used to detect the expression level of miR-503 in melanoma tissues and cell lines; miR-503 mimics or miR-503 inhibitors were used to perform transient transfection to A375 cells, and Transwell assay was used to test cell migration and invasion abilities. The potential binding site of miR-503 on the CXCL9 was predicted by using online bioinformative databases, and the result was verified by double luciferase assay. The migration and invasive ability of A375 cells were detected after transfected with CXCL9 alone or in combination with miR-503. Results: Expression of miR-503 was significantly downregulated in melanoma tissues when compared with adjacent normal tissues (t=6.602, P=0.000). In addition, expression of miR-503 in the three melanoma cell lines (A375, SK-MEL-1 and SK-MEL-5) was significantly lower than that in the HEM cells (F=19.445, P=0.000). Migration and invasion abilities in A375 cells after transfection of miR-503 mimics were significantly inhibited (P=0.017, P=0.049), while in A375 cells after transfection of miR-503 inhibitors were promoted (P=0.004, P=0.000). CXCL9 was the direct target gene of miR-503 and promoted the migration and invasion of A375 cells (P=0.000, P=0.009), of which effectiveness was able to be reversed by miR-503. Conclusion: The miR-503 suppresses migration and invasion of melanoma cells by targeted-regulating CXCL9.