Fascin-1对恶性黑色素瘤细胞增殖和凋亡的影响及其作用机制

发布时间：2019-02-16 05:12

【摘要】：目的探讨Fascin-1对恶性黑色素瘤细胞增殖和凋亡的影响及其作用机制。方法体外培养人恶性黑色素瘤细胞A375和人皮肤成纤维细胞CCC-HSF-1。将A375细胞随机分为CON组和Fascin-1-shRNA组,分别按照10 MOI的感染复数加入CON和Fascin-1-shRNA病毒,培养48 h。采用qRT-PCR法及Western blotting法检测常规培养的CCC-HSF-1、A375细胞以及CON组、Fascin-1-shRNA组Fascin-1 mRNA和蛋白表达。分别采用Dye67染色法、Annexin V/PI染色法检测CON组、Fascin-1-shRNA组细胞增殖指数及凋亡率,采用Western blotting法检测Bcl-2、Bax、Bad蛋白表达。结果常规培养的CCC-HSF-1、A375细胞中Fascin-1 mRNA相对表达量分别为0.21±0.015、1.00±0.01,Fascin-1蛋白相对表达量分别为0.12±0.04、0.75±0.11,二者比较P均0.01。CON组、Fascin-1-shRNA组Fascin-1 mRNA相对表达量分别为1.00±0.01、0.19±0.04,Fascin-1蛋白相对表达量分别为1.28±0.17、0.24±0.10,两组比较P均0.01。Fascin-1-shRNA组培养3、4天时细胞增殖指数均低于CON组同期(P均0.05)。Fascin-1-shRNA组细胞凋亡率高于CON组(P0.01)。Fascin-1-shRNA组Bcl-2蛋白相对表达量低于CON组,Bax、Bad蛋白相对表达量高于CON组(P均0.01)。结论 Fascin-1表达下调可抑制恶性黑色素瘤细胞增殖并诱导其凋亡,其机制可能与下调细胞中Bcl-2表达及上调Bax、Bad表达有关。
[Abstract]:Objective to investigate the effect and mechanism of Fascin-1 on proliferation and apoptosis of malignant melanoma cells. Methods Human malignant melanoma cell line A375 and human skin fibroblast CCC-HSF-1. were cultured in vitro. A375 cells were randomly divided into CON group and Fascin-1-shRNA group. According to the complex number of 10 MOI infection, CON and Fascin-1-shRNA virus were added to A375 cells for 48 h. QRT-PCR and Western blotting were used to detect the expression of Fascin-1 mRNA and protein in cultured CCC-HSF-1,A375 cells, CON group and Fascin-1-shRNA group. The cell proliferation index and apoptosis rate of CON group and Fascin-1-shRNA group were detected by Dye67 staining and Annexin V/PI staining respectively. Bcl-2,Bax,Bad protein expression was detected by Western blotting method. Results the relative expression of Fascin-1 mRNA in CCC-HSF-1,A375 cells was 0.21 卤0.0151.00 卤0.01Fascin-1, 0.12 卤0.04 卤0.75 卤0.11, respectively. The relative expression of Fascin-1 mRNA in Fascin-1-shRNA group was 1.00 卤0.01U 0.19 卤0.04Fascin-1 protein, respectively 1.28 卤0.170.24 卤0.10, respectively. The cell proliferation index of 0.01.Fascin-1-shRNA group was lower than that of CON group after 3 days of culture (P0. 05). The apoptosis rate of Fascin-1-shRNA group was higher than that of CON group (P0. 01). The Bcl of Fascin-1-shRNA group was higher than that of CON group (P0. 01). The relative expression of -2 protein in CON group was lower than that in CON group. The relative expression of Bax,Bad protein was higher than that of CON group (P 0.01). Conclusion down-regulation of Fascin-1 expression can inhibit the proliferation and induce apoptosis of malignant melanoma cells, which may be related to down-regulation of Bcl-2 expression and up-regulation of Bax,Bad expression.
【作者单位】：武汉市第一医院;华中科技大学同济医学院附属武汉中心医院;
【分类号】：R739.5

【参考文献】