摘要
目的:观察CXCR4基因沉默后人恶性黑素瘤A375细胞增殖及侵袭能力变化。方法:体外培养人恶性黑素瘤A375细胞,分为A、B、C 3组,分别为siRNA组,转染空病毒载体的阴性对照组,细胞不做任何处理的空白对照组。转染后1、3、5、7d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,采用流式细胞分析术,Anexin-ⅴ-PI双染色检测细胞早期凋亡的变化。转染后7d采用RT-PCR检测CXCR4siRNA表达情况。结果:转染后C组细胞出现不同程度的皱缩、破裂,漂浮细胞增多。转染后1、3、5、7d,A组OD值低于B、C组(P<0.05),A组早期凋亡率的变化与B、C组有统计学差异(P<0.05)。RT-PCR半定量分析结果显示,A、B、C组CXCR4基因mRNA校正值分别为(0.149±0.043),(0.215±0.030),(0.221±0.020),A组与B、C组相比差异有统计学意义(P<0.05)。结论:CXCR4基因沉默后A375细胞增殖被抑制,早期凋亡增加,CXCR4基因成为恶性黑素瘤治疗的新靶点。
Objective：To explore the proliferation and apoptosis of A375 cells after silencing CXCR4 gene.Methods：Human malignant melanoma A375 cells were cultivated and divided into groups A,B and C：siRNA group with adenovirus carrying RNA interference,negative group with vacant adenovirous vector,and control group without any infection.The differentiation and morphology of cells were observed by inverted microscope.1, 3, 5, 7days after transfection,cells proliferation ability was determined by MTT,and apoptosis was detected by Annexin V-FITC/PI.The expressions of CXCR4 mRNA was detected by RT-PCR 7 days after transfection.Results：The cells proliferation ability of group A at 1, 3, 5 and 7days after transfection was lower than those in groups B and C（P〈0.005）.The expression of CXCR4 siRNA decreased in the A group than groups B and C.CXCR4 gene silenced by siRNA could inhibit the growth of A375 cell proliferation（P〈0.05）and induce early cells apoptosis（P〈0.05）.Conclusion：The cells proliferation ability of A375 is inhibited and apoptosis is induced after silencing CXCR4 gene.
出处
《陕西医学杂志》
CAS
2017年第4期411-413,共3页
Shaanxi Medical Journal
基金
国家自然科学基金资助项目(30440086)
