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目的探讨槲皮素对人食管癌Eca-109细胞的分化诱导作用。方法将培养的Eca-109细胞分为两组:①加槲皮素组(Q组);②不加药物对照组(C组)。同时培养48h,收集各组细胞制备成细胞硝酸纤维素膜(NCM)标本及提取RNA制备RNA硝酸纤维素膜标本。分别对细胞硝酸纤维素膜(NCM)标本进行PCNA,VEGF的免疫斑点印迹阵列;对RNA硝酸纤维素膜标本EGFR,c-myc及wtp53的RNA斑点印迹阵列。印迹阵列皆用岛津薄层色谱扫描仪(TLC)进行波长523nm的扫描数值比较。结果①免疫斑点印迹显示:Q组的PCNA-IR,VEGF-IR的TLC扫描数值分别比C组低5.5倍,3.5倍;②RNA斑点印迹阵列显示,与C组相比:Q组的EGFR,c-myc的RNA斑点TLC扫描数值分别下调了7倍,2.2倍。而wtp53的RNA斑点TLC扫描数值则上调了2.2倍。结论槲皮素可下调作为肿瘤细胞恶性程度生物学指标的PCNA,EGFR,VEGF,c-myc等的表达,上调抑癌基因wtp53的表达,表明对人食管癌Eca-109细胞具有一定的诱导分化效应。
Objective To investigate the differentiation-inducing effect of quercetin on human esophageal carcinoma Eca-109 cells. Methods The cultured Eca-109 cells were divided into two groups: ① Quercetin group (Q group); ② No drug control group (C group). At the same time, the cells were cultured for 48 hours. The cells were collected to prepare the cell nitrocellulose membrane (NCM) specimens and the RNA was extracted to prepare the RNA nitrocellulose membrane specimens. The immunofluorescent dot blot arrays of PCNA and VEGF were respectively prepared on NCM specimens and the RNA dot blot arrays of EGFR, c-myc and wtp53 on RNA-nitrocellulose membrane specimens. The imprinted arrays were compared by scanning at 523 nm using a Shimadzu Thin Layer Chromatography Scanner (TLC). Results ① Immunofluorescent dot blot showed that the values of TLC in PCNA-IR and VEGF-IR in Q group were 5.5 times and 3.5 times lower than those in group C respectively. (2) Compared with group C, -myc RNA dot TLC scan values were down 7 times, 2.2 times. The wtp53 RNA spot TLC scans increased 2.2-fold. Conclusions Quercetin can down-regulate the expression of PCNA, EGFR, VEGF, c-myc and other genes which are the biological indicators of malignant degree of tumor cells, upregulate the expression of the suppressor gene wtp53, indicating that it has certain induction and differentiation on human esophageal cancer Eca-109 cells effect.