国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2015年
6期
401-405
,共5页
牛蒡子苷元%凋亡%化疗敏感性%顺铂%肺癌
牛蒡子苷元%凋亡%化療敏感性%順鉑%肺癌
우방자감원%조망%화료민감성%순박%폐암
Arctigenin%Apoptosis%Chemosensitization%Cisplatin%Lung cancer
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Survivin cDNA,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24 h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.
目的 研究Survivin基因在牛蒡子苷元促進肺癌細胞化療敏感性中的作用.方法 應用RT-PCR技術從H460細胞中擴增全長人Survivin cDNA,併將其剋隆至pcDNA3.1(+)錶達載體,構建人Survivin基因真覈錶達質粒.將pcDNA3.1-Survivin重組質粒或空載體分彆轉染H460細胞,孵育24 h後,應用Western blot法檢測Survivin過錶達情況;轉染細胞與牛蒡子苷元單獨或聯閤順鉑處理,應用Annexin-Ⅴ/PI染色法檢測細胞凋亡情況.結果 與未轉染和轉染空載體質粒的H460細胞相比,轉染pcDNA3.1-Survivin重組質粒H460細胞Survivin水平顯著增加,提高大約10倍.低劑量順鉑對H460細胞Survivin蛋白錶達影響不顯著,而牛蒡子苷元可顯著抑製Survivin蛋白錶達水平.噹牛蒡子苷元與順鉑聯閤作用時,Survivin蛋白錶達抑製程度更大.與轉染空載體質粒相比,轉染Survivin錶達重組質粒的細胞對牛蒡子苷元單獨或聯閤順鉑誘導的凋亡具有抗性,細胞凋亡比率顯著下降.結論 牛蒡子苷元抑製人肺癌細胞中Survivin基因錶達.牛蒡子苷元促人肺癌細胞順鉑敏感性與抑製Survivin通路有關.
목적 연구Survivin기인재우방자감원촉진폐암세포화료민감성중적작용.방법 응용RT-PCR기술종H460세포중확증전장인Survivin cDNA,병장기극륭지pcDNA3.1(+)표체재체,구건인Survivin기인진핵표체질립.장pcDNA3.1-Survivin중조질립혹공재체분별전염H460세포,부육24 h후,응용Western blot법검측Survivin과표체정황;전염세포여우방자감원단독혹연합순박처리,응용Annexin-Ⅴ/PI염색법검측세포조망정황.결과 여미전염화전염공재체질립적H460세포상비,전염pcDNA3.1-Survivin중조질립H460세포Survivin수평현저증가,제고대약10배.저제량순박대H460세포Survivin단백표체영향불현저,이우방자감원가현저억제Survivin단백표체수평.당우방자감원여순박연합작용시,Survivin단백표체억제정도경대.여전염공재체질립상비,전염Survivin표체중조질립적세포대우방자감원단독혹연합순박유도적조망구유항성,세포조망비솔현저하강.결론 우방자감원억제인폐암세포중Survivin기인표체.우방자감원촉인폐암세포순박민감성여억제Survivin통로유관.
Objective To explore the role of survivin in the chemosensitization of arctigenin in lung cancer H460 cells.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-transfected cells had a 10-fold elevation in the survivin expression.Low-dose cisplatin did not significantly affect the expression of survivin in H460 cells.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.
