转染c-kit基因对A375细胞增殖和凋亡的影响

摘要

目的:明确转染c-kit基因对A375细胞增殖、凋亡的影响.方法:体外培养人恶性黑素瘤A375细胞,分为A组(实验组)、B组(转染空病毒载体的阴性对照组)、C组(空白对照组).转染后采用RT-PCR检测c-kit mRNA表达情况,在倒置相差显微镜观察A375细胞分化及形态,MTT比色分析法检测对A375细胞增殖抑制的影响,流式细胞分析术,Anexin-v-PI双染色检测细胞早期凋亡的变化.结果:转染后A组细胞出现不同程度的皱缩、破裂,漂浮细胞增多.RT-PCR半定量分析结果显示,A组扩增出c-kit cDNA 230 bp大小片段,而B、C组则没有扩增出相应大小的片段.转染后A组OD值低于B、C组(P<0.05),A组早期凋亡率高于B、C组有显著性差异(P<0.05).结论:转染c-kit后A375细胞增殖被抑制,早期凋亡增加.%Objective: To determine the effect of c-kit gene on the proliferation and apoptosis of A375 cells.Methods: Human malignant melanoma A375 cells were cultured and divided into the group A (adenovirous vector group), group B (vacant adenovirous vector group) and group C (blank control group).The expression level of c-kit mRNA was detected by RT-PCR.The differentiation and morphology of the cells were observed under the inverted microscope.The proliferation ability and apoptosis of A375 cells were detected by MTT and Annexin V-FITC/PI.Results: There was more shrinking and splinter cells in group A than those in group B and C.The c-kit cDNA 230bp was detected in group A but not in group B and C.The proliferation ability of A375 cells in group A was lower than those in group B and C, with a significant difference (P<0.005).The apoptosis rate of A375 cells in the group A was higher than that in the group B and C (P<0.05).Conclusion: The proliferation ability of A375 cells is inhibited and apoptosis is induced after transfected by c-kit gene.