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目的观察靶向survivin的RNA干扰对人恶性黑色素瘤A375细胞的影响。方法将处于对数生长期的黑色素瘤A375细胞随机分为3组:阳性组、阴性对照组、空白对照组,将设计并构建好的survivin-siRNA慢病毒载体分别转染入各组细胞内,实时定量PCR及Western-blot法分别检测转染前后3组细胞survivin mRNA及蛋白的表达情况。结果成功设计并构建了survivin-siRNA慢病毒载体,与空白对照组相比,转染survivin-siRNA的阳性组survivin mRNA及蛋白的表达明显降低,抑制率分别为60%、87.38%,差异有统计学意义(P<0.05)。结论靶向survivin的siRNA能够明显抑制恶性黑色素瘤A375细胞的survivin基因的表达。
Objective To observe the effect of RNAi targeting survivin on human malignant melanoma A375 cells. Methods Melanoma A375 cells in logarithmic growth phase were randomly divided into three groups: positive group, negative control group and blank control group. The designed and constructed survivin-siRNA lentiviral vector were transfected into each group of cells, Real-time quantitative PCR and Western-blot were used to detect the expression of survivin mRNA and protein in 3 groups before and after transfection. Results The survivin-siRNA lentiviral vector was successfully designed and constructed. Compared with the blank control group, the expression of survivin mRNA and protein in the transfected survivin-siRNA group was significantly reduced, with the inhibition rates of 60% and 87.38%, respectively Significance (P <0.05). Conclusion siRNA targeting survivin can significantly inhibit the expression of survivin gene in human melanoma A375 cells.