环状RNA Circ_0001073在恶性黑素瘤中的表达及其对A375细胞恶性表型的影响

目的:通过检测环状RNA Circ_0001073在恶性黑素瘤细胞与正常表皮黑素细胞中的表达差异,及其对恶性黑素瘤A375细胞恶性表型的影响,阐明Circ_001073在恶性黑素瘤细胞中的表达及功能。方法:采用聚合酶链式反应(Real-time quantitative polymerase chain reaction, RT-QPCR)方法检测正常表皮黑素细胞和恶性黑素瘤细胞中Circ_0001073的表达水平;应用小干扰RNAs沉默恶性黑素瘤A375细胞中Circ_0001073表达,5-乙炔基-2'-脱氧尿苷(5-Ethynyl-2'-deoxyuridine,EdU)实验检测沉默Circ_0001073对A375细胞增殖的影响,平板克隆实验检测沉默Circ_0001073对A375细胞克隆形成的影响,细胞划痕实验评估沉默Circ_0001073对A375细胞迁移能力的影响,Transwell实验评估沉默Circ_0001073对A375细胞侵袭能力的影响。结果:qRT-PCR结果显示Circ_0001073在恶性黑素瘤细胞中的表达低于正常表皮黑素HEMa-LP细胞(P0.05)。应用Circ_0001073si RNA转染A375细胞后,Circ_0001073的表达水平较转染Circ_0001073 NC的细胞显著降低(P0.05)。EdU实验结果显示:沉默A375细胞中Circ_0001073的表达后,A375细胞增殖能力显著增强(P0.05);平板克隆实验结果显示:沉默A375细胞中Circ_0001073的表达后,A375细胞克隆形成能力显著增强(P0.05);细胞划痕实验结果显示:沉默A375细胞中Circ_0001073的表达后,细胞迁移能力显著增强(P0.05);Transwell实验结果显示:沉默A375细胞中Circ_0001073的表达后,细胞侵袭能力显著增强(P0.05)。结论:Circ_0001073在恶性黑素瘤细胞中低表达,并可抑制A375细胞的增殖、克隆形成、迁移及侵袭。

Expression of Circular RNA Circ_0001073 in Malignant Melanoma and Its Effects on Malignant Phenotypes of A375 Cells

ABSTRACT Objective: To demonstrate the expression and function of Circ_0001073 in A375 cells by investigating the expression of circular RNA Circ_0001073 in malignant melanoma cells and normal epidermal melanoma cells, and exploring the effects of Circ_0001073 on malignant phenotypes of A375 cells. Methods: Real-time quantitative polymerase chain reaction (RT-QPCR) was performed to detect the expression of Circ_0001073 in normal epidermal melanoma cells and malignant melanoma cells. Silencing the expression of Circ_0001073 in A375 cells by small interfering RNAs (siRNAs). 5-Ethynyl-2''-deoxyuridine (EdU) assays were performed to detect the effects of Circ_0001073 knockdown on cell proliferation of A375 cells; Colony forming assays were performed to detect the effects of Circ_0001073 knockdown on colony forming ability of A375 cells; Wound healing assays were perform to detect the effects of Circ_0001073 knockdown on migrative ability of A375 cells; Transwell assays were performed to detect the effects of Circ_0001073 knockdown on invasive ability of A375 cells. Results: Compared to normal epidermal melanocytes HEMa-LP cells, Circ_0001073 was downregulated in malignant melanoma cells(P<0.05). After silencing the expression of Circ_0001073 by siRNAs in A375 cells, the expression of Circ_0001073 was significantly down-regulated in A375 cells transfected with Circ_0001073 siRNA compared to A375 cells transfected with Circ_0001073 NC(P<0.05). EdU assays indicated the cell proliferation was enhanced after silencing the expression of Circ_0001073 in A375 cells(P<0.05); Colony forming assays indicated the colony forming ability was enhanced after silencing the expression of Circ_0001073 in A375 cells (P<0.05); Wound healing assays indicated the cell migrative ability was enhanced after silencing the expression of Circ_0001073 in A375 cells (P<0.05); Transwell assays indicated the cell invasive ability was enhanced after silencing the expression of Circ_0001073 in A375 cells(P<0.05). Conclusion: Circ_0001073 is down-regulated in malignant melanoma cells, and Circ_0001073 could inhibits cell proliferation, colony forming, migrative and invasive ability of A375 cells.