Survivin基因在牛蒡子苷元增强人肺癌H460细胞化疗敏感性中的作用

Biological functions of Survivin in chemosensitization of arctigenin in lung cancer H460 cells

目的研究Survivin基因在牛蒡子苷元促进肺癌细胞化疗敏感性中的作用。方法应用RT-PCR技术从H460细胞中扩增全长人SurvivincDNA，并将其克隆至pcDNA3．1（＋）表达载体，构建人Survivin基因真核表达质粒。将pcDNA3．1-Survivin重组质粒或空载体分别转染H460细胞，孵育24h后，应用Western blot法检测Survivin过表达情况；转染细胞与牛蒡子苷元单独或联合顺铂处理，应用Annexin-V／PI染色法检测细胞凋亡情况。结果与未转染和转染空载体质粒的H460细胞相比，转染pcDNA3．1-Survivin重组质粒H460细胞Survivin水平显著增加，提高大约10倍。低剂量顺铂对H460细胞Survivin蛋白表达影响不显著，而牛蒡子苷元可显著抑制Survivin蛋白表达水平。当牛蒡子苷元与顺铂联合作用时，Survivin蛋白表达抑制程度更大。与转染空载体质粒相比，转染Survivin表达重组质粒的细胞对牛蒡子苷元单独或联合顺铂诱导的凋亡具有抗性，细胞凋亡比率显著下降。结论牛蒡子苷元抑制人肺癌细胞中Survivin基因表达。牛蒡子苷元促人肺癌细胞顺铂敏感性与抑制Survivin通路有关。

Objective To explore the role of survivin in the chemosensitization of arctigenin in lung cancer H460 ceils. Methods Human full-length Survivin cDNA was amplified by polymerase chain reaction and cloned into pcDNA3.1 （＋） expression vector. The resultant survivin-expressing plasmid or empty vector was individually transfected into H460 cells using Lipofectamine 2000. Twenty-four hours after transfection,survivin expression in H460 cells was tested by Western blot. Transfected cells were exposed to arctigenin alone or combined with cisplatin and cell apoptosis was assessed using Annexin-V/ PI staining methods. Results Compared to untransfected and empty vector-transfected H460 cells, survivin-transfeeted cells had a 10-fold elevation in the survivin expression. Low-dose cisplatin did not significantly affect the expression of survivin in H460 ceils. In contrast, arctigenin significantly inhibited the expression of survivin protein. When cisplatin was combined with arctigenin, a greater inhibition of survivin expression was observed. Compared to transfection of empty vector, pre-transfection with pcDNA3. 1-Survivin plasmid significantly reversed the pro-apoptotic activity of arctigenin alone or combined with cisplatin,leading to a significant reduction in the apoptotic index. Conclusions Arctigenin can inhibit the expression of survivin in lung cancer H460 cells. Arctigenin-mediated chemosensitization of lung cancer H460 cells to cisplatin is associated with suppression of survivin signaling pathway. [