外源性CSMD1对黑色素瘤A375细胞活性的影响

Effects of exogenous CSMD1 on activity of A375 melanoma cells

目的 研究经pCDNA-3.0-CSMD1转染的A375细胞的活性,探讨外源性CSMD1的表达对黑色素瘤细胞活性的影响.方法 培养人黑色素瘤A375细胞株,利用前期实验获得的CSMD1基因,构建质粒、转染细胞.A375细胞分为：A组,未转染的A375细胞;B组,经pCDNA-3.0转染的A375细胞;C组,经pCDNA-3.0-CSMD1转染的A375细胞.应用MTT法、细胞周期分析法、细胞凋亡相关检测（DAPI染色、Annexin V-FITC/PI双染、Transwell细胞迁移实验）等方法,检测A375细胞的活性,并进行统计学分析.结果 C组细胞的增殖率明显低于其他两组细胞（P＜0.05）;C组细胞G1期的比例高于其他两组细胞（P＜0.05）;仅在C组细胞中发现凋亡细胞核固缩,并在48 h内出现核碎裂;C组细胞凋亡率为32.7％,明显高于A组（6.6％）和B组（8.4％）细胞（P＜0.05）;与A组和B组细胞相比,C组细胞极少迁移至聚碳酸酯膜下层（P＜0.05）.结论 外源性CSMD1可以抑制黑色素瘤A375细胞的活性,这有可能为黑色素瘤的治疗提供一种新的途径.

Objective To study the activity of pCDNA-3.0-CSMD1 transfected A375 cells,and investigate effects of exogenous CSMD1 on activity of A375 melanoma cells.Methods The cultured A375 melanoma cells and CSMD1 gene which obtained by prevenient experiments were used to construct the plasmid and transfected A375 cells.The A375 cells were divided into the A group （the cells without transfection）,the B group （the cells transfected with pCDNA-3.0） and the C group （the cells transfected with pCDNA-3.0 and CSMD1）.The activity of A375 cells was detected with MTT assay,cell cycle analysis,measurement of apoptotic cell death （DAPI staining assty,Annexin-V FITC/PI double staining assay,Transwell migration assay）and the statistical analysis aws also performed.Results The rate of cell proliferation in the C group was lower than those in the other groups （P＜ 0.05）,and its proportion of G1 phase was higher （P＜ 0.05）.Apoptotic nuclear condensation with nuclear fragmentation was only observed in the C group within 48 hours.The percentage of apoptosis in the C group was 32.7 ％,which was significantly higher compared to Group A （6.6 ％） and Group B （8.4％）,P＜0.05.The cells in the C group significantly less migrated to the lower membrane compared to cells in the A and B groups （P ＜ 0.05）.Conclusion Exogenous CSDM1 can inhibit activity of melanoma A375 cells,and may provide a novel target for clinical treatment.