目的:研究理冲汤对体外培养人子宫肌瘤细胞p53信号通路基因表达谱的影响。方法:体外原代培养子宫肌瘤细胞并传代后,加入理冲汤含药血清,继续培养。选用功能分类PCR p53信号通路基因芯片(PCR p53 Signaling PathwayMicroarray),从空白对照组、理冲汤组子宫肌瘤细胞中进行样品的RNA抽提及纯化,采用紫外吸收测定法及变性琼脂糖凝胶电泳进行RNA质量检测,合成cDNA,最后进行实时定量PCR。结果:所采用的PCR p53信号通路基因芯片共检测96条基因,理冲汤组与空白对照组比较,共17条(17.7%)基因表达发生明显变化,其中10条基因表达量明显下降(Fold Down-Regulation〈-2),7条基因表达量上升(Fold Up-Regulation〉2)。按功能将其进行大体分类,涉及到细胞凋亡、细胞周期、细胞生长增殖和分化、DNA修复等多方面。结论:理冲汤对子宫肌瘤p53信号通路的细胞凋亡、细胞周期、细胞生长增殖和分化、DNA修复等相关基因均有调节作用,提示细胞内外的信号传导是子宫肌瘤发病的关键环节,理冲汤对子宫肌瘤p53信号通路相关基因表达具有调控作用。
Objective:To study the effect of Lichong decoction on p53 signaling pathway gene expression profile in human uterine leiomyoma cells in vitro.Method: The primary cultured uterine leiomyoma cells were treated with Lichong decoction.Using PCR p53 signaling pathway microarray,all RNA was gained and cleanuped from human uterine leiomyoma cells in blank control group and Lichong decoction group.RNA yield and quality were assessed by UV absorbance and denaturing agarose gel electrophoresis,first strand cDNA Synthesis,performing Real-Time PCR finally.Result: Lichong decoction group was compared with blank control group among 96 pieces genes in PCR p53 signaling pathway microarray,17 in total(17.7%)genes expression had obviously changed,10 pieces went down obviously(Fold Down-Regulation-2),7 piece significantly went up(Fold Up-Regulation2).The genes were generally classified,according to their functions,into apoptosis,cell cycle,cell growth,proliferation and differentiation and DNA repair.Conclusion: Lichong decoction can regulate correlation genes of apoptosis,cell cycle,cell growth,proliferation and differentiation and DNA repair in p53 signaling pathway of uterine leiomyoma,whichshow that signal-transmit between in and out of cell is the key link in uterine leiomyoma process,Lichong decoction has the effect of regulation and control of p53 signaling pathway gene expression profile.