纳米雄黄对皮肤鳞癌A431细胞p53、Bcl-2表达的影响及相关机制探讨

Investigation of the effect and mechanism of realgar nano on the expression of P53 and Bcl- 2 in skin squamous cell carcinoma A431 cells

目的探讨纳米雄黄对皮肤鳞癌A431细胞p53、Bcl-2表达的影响及相关机制。方法通过细胞培养,应用逆转录-聚合酶链反应(RT-PCR)技术、免疫组化法观察纳米雄黄干预p53、Bcl-2在人皮肤鳞癌A431细胞中的表达情况。结果各浓度纳米雄黄干预组、注射用顺铂(DDP)干预组及联合用药组的Bcl-2阳性率均明显低于对照组(P<0.05);随着纳米雄黄浓度的升高,Bcl-2阳性率也明显降低(P<0.05);联合用药组的Bcl-2阳性率明显低于各浓度纳米雄黄干预组和DDP干预组(P<0.05)。各浓度纳米雄黄干预组、DDP干预组及联合用药组的p53的相对灰度值(RATIO)均明显高于对照组(P<0.05);随着纳米雄黄浓度的升高,p53的RATIO明显升高(P<0.05);联合用药组的p53的RATIO明显高于各浓度纳米雄黄干预组和DDP干预组(P<0.05)。各浓度纳米雄黄干预组条带灰度高于对照组;5%、10%、20%纳米雄黄干预组条带亮度依次变大;联合用药组条带亮度较DDP干预组、各浓度纳米雄黄干预组大,内参β-actin见不到肉眼可见的浓度变化。结论纳米雄黄可上调p53表达和下调Bcl-2表达,与DDP联合应用有协同作用,可能是纳米雄黄诱导人皮肤鳞癌A431细胞凋亡的机制之一。

Objective To investigate the effect and mechanism of realgar nano on the expression of P 53 and Bcl-2 in skin squamous cell carcinoma A 431 cells.Methods The effect of realgar nano on the expression of P 53 and Bcl-2 in skin squamous cell carcinoma A 431 cells were observed by cell culture ,RT-PCR and immunohisto-chemistry.Results Positive rate of Bcl-2 in different concentration groups of realgar nano ,DDP group and combi-nation drug therapy group was higher than that in control group （P〈0.05）.Positive rate of Bcl -2 was decreased with the increase of realgar nano concentration （P〈0.05）.Positive rate of Bcl-2 in combination drug therapy group was lower than that in different concentration groups of realgar nano and DDP group （P〈0.05）.The mean gray value of p53 in different concentration groups of realgar nano ,DDP group and combination drug therapy group was higher than that in control group（P〈0.05）.The mean gray value of p53 was increased with the increase of realgar nano concen-tration（P〈0.05）.The mean gray value of p53 in combination drug therapy group was higher than that in different concentration groups of realgar nano and DDP group （P〈0.05）.Striped gray in different concentration groups of real-gar nano was higher than that in control group （P〈0.05）.Striped gray in 5%,10%and 20%realgar nano group was increased.Striped gray in combination drug therapy group was higher than that in different concentration groups of re -algar nano and DDP group（P〈0.05）.Conclusion Realgar nano may down regulate the expression of Bcl -2 and has synergistic effect by combination of DDP ,one of the mechanisms may be nano realgar -induced human skin squa-mous cell apoptosis of A431 cells.