circFNDC3B通过抑制miR147表达促进恶性黑素瘤迁移与侵袭
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国家自然科学基金项目(81802935)


circFNDC3B promotes cell migration and invasion via inhibiting miR147 expression in malignant melanoma cells
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    摘要:

    【摘要】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HEMaLP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siNC和sicircFNDC3B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Ecadherin和Ncadherin表达,应用RTQPCR进行检则miRNAs表达。结果 与正常表皮黑素细胞HEMaLP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),Ecadherin表达显著增强,Ncadherin表达显著减弱,抑癌性miRNA miR147表达升高。结论 circFNDC3B在恶性黑素瘤细胞中高表达,可通过抑制miR147表达促进上皮间质转化,进而促进细胞迁移及侵袭。

    Abstract:

    【Abstract】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. 

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  • 在线发布日期: 2020-11-11
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