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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 Li Chong Decoction on the gene expression profile of p53 signaling pathway in human uterine fibroids. Methods: Primary uterine fibroids were cultured in vitro and passaged, and the drug-containing serum from Ritong decoction was added to continue the culture. The functional classification PCR p53 Signaling Pathway Microarray was used to extract and purify the RNA from the uterine leiomyoma cells of the blank control group and the Li Chong Tang group. The UV absorbance assay and denaturing Sepharose Electrophoresis RNA quality testing, synthesis of cDNA, the final real-time quantitative PCR. Results: A total of 96 genes were detected by the PCR p53 signal pathway gene chip, 17 genes (17.7%) were significantly changed in Li Chong Tang group compared with the blank control group, of which 10 genes were significantly decreased (Fold Down-Regulation <-2), the expression of seven genes increased (Fold Up-Regulation> 2). According to the function of its general classification, related to apoptosis, cell cycle, proliferation and differentiation of cell growth, DNA repair and many other aspects. CONCLUSION: Li Chong Decoction regulates the apoptosis, cell cycle, proliferation and differentiation, DNA repair and other related genes of p53 signaling pathway in uterine fibroids, suggesting that intracellular and extracellular signal transduction is the key link in the pathogenesis of uterine fibroids , Red soup on uterine fibroids p53 signaling pathway related gene expression regulation.