靶向沉默Zwilch对人皮肤恶性黑素瘤细胞增殖及凋亡的影响

Effects of targeted silencing Zwilch on proliferation and apoptosis in human cutaneous malignant melanoma cell

目的探讨靶向沉默Zwilch对人皮肤恶性黑素瘤细胞增殖及凋亡的影响。方法通过感染shRNA慢病毒靶向沉默A375细胞Zwilch的表达及作用,实时荧光定量PCR及Western blotting检测其沉默效果;采用CCK-8和Annexin V/PI染色分别检测靶向沉默Zwilch对A375细胞的增殖及凋亡的影响。结果慢病毒感染72 h后,sh-Zwilch细胞的Zwilch mRNA及蛋白表达水平均显著下降(P<0.05),shRNA慢病毒靶向沉默A375中Zwilch的表达和作用与空白组比较,sh-Zwilch组细胞活性随时间显著下降(P<0.05),凋亡比例明显上升(P<0.05)。结论靶向沉默Zwilch能够有效抑制人皮肤恶性黑素瘤细胞增殖并诱导其凋亡,从而有望成为其基因靶向治疗的一种新靶点和新策略。

Objective To investigate the effects of targeted silencing Zwilch on proliferation and apoptosis in human cutaneous malignant melanoma cell.MethodsTargeted silence the expression and effect of Zwilch(sh-Zwilch)in A375 cells was examined by shRNA lentivirus infected.Then the expression levels of Zwilch mRNA and protein in the cell were detected by real-time quantitative PCR and western blotting respectively.The cellular viability was analyzed by CCK-8 assay.The apoptosis was evaluated by Annexin V/PI staining.ResultsCompared with the control cell,A375 cell infected with sh-Zwilch showed significant decrease of expression level of Zwilch mRNA and protein.Subsequently,Zwilch targeted silencing significantly suppressed the cellular viability in a time-dependent manner and increased the apoptosis rate in A375 cell(P<0.05).ConclusionTargeted silence of Zwilch could inhibit the proliferation and promotes the apoptosis of A375 cell,and Zwilch maybe a potential target for human CMM gene therapy.