circFNDC3B通过抑制miR-147表达促进恶性黑素瘤迁移与侵袭

circFNDC3B promotes cell migration and invasion via inhibiting miR-147 expression in malignant melanoma cells

目的探究正常表皮黑素细胞和恶性黑素瘤细胞中环状RNA circFNDC3B的表达水平及其对恶性黑素瘤A2058细胞迁移和侵袭的影响。方法RT-QPCR实验检测恶性黑素瘤A2058和A375细胞和正常表皮黑素HEMa-LP细胞中circFNDC3B表达水平;siRNAs敲低恶性黑素瘤A2058细胞中circFNDC3B的表达;转染si-NC和si-circFNDC3B的A2058细胞分别为对照组和实验组,应用细胞划痕实验比较细胞迁移距离,应用免疫印迹实验检测E-cadherin和N-cadherin表达,应用RT-QPCR进行检则miRNAs表达。结果与正常表皮黑素细胞HEMa-LP相比,circFNDC3B在恶性黑素瘤细胞中的表达上调。沉默A2058细胞中circFNDC3B表达后,细胞的迁移和侵袭能力均显著降低(P<0.05),E-cadherin表达显著增强,N-cadherin表达显著减弱,抑癌性miRNA miR-147表达升高。结论circFNDC3B在恶性黑素瘤细胞中高表达,可通过抑制miR-147表达促进上皮间质转化,进而促进细胞迁移及侵袭。

Objective To investigate the expression of circular RNA_circFNDC3B in normal epidermal melanoma cells and malignant melanoma cells and explore the effects of circFNDC3B on migration and invasion of A2058 cells.Methods The expression of circFNDC3B and in malignant melanoma cells and normal epidermal melanoma cells was evaluated by real-time quantitative polymerase chain reaction(RT-QPCR).Small interfering RNAs(siRNAs)were used to knockdown the expression of circFNDC3B in A2058 cells.Migrative distance of A2058 cells transfected with si-NC or si-circFNDC3B was measured by wound healing assays.Invasive ability of A2058 cells transfected with si-NC or si-circFNDC3B was measured by transwell assays.Expression of E-cadherin and N-cadherin in A2058 cells transfected with si-NC or si-circFNDC3B was measure by western blot.Expression of microRNAs(miRNAs)in A2058 cells transfected with si-NC or si-circFNDC3B was measure by RT-QPCR.Results CircFNDC3B was abnormally higher in malignant melanoma cells than that in epidermal melanocytes HEMa-LP cells.Silencing the expression of circFNDC3B in A2058 cells resulted in significantly decreased abilities of cell migration and invasion(P<0.05).The expression of E-cadherin which was the epithelial cells marker was enhanced.The expression of N-cadherin which was the mesenchymal cells marker was suppressed.miR-147 expression which was an established tumor suppressive miRNA was upregulated.Conclusion CircFNDC3B is up-regulated in malignant melanoma cells,and promotes cell migration and invasion of A2058 cells and epithelial-mesenchymal transition(EMT)via inhibiting miR-147 expression.