吗啡对人胃癌MGC-803细胞p53 mRNA和E2F-1 mRNA表达的影响
Effect of morphine on expression of p53 mRNA and E2F-1 mRNA in human gastric carcinoma cell line MGC-803
摘要目的 探讨吗啡对人胃癌MGC-803细胞p53 mRNA和E2F-1 mRNA表达的影响.方法 人胃癌MGC-803细胞以1×103/ml或2×105/ml密度接种于6孔培养板中,1 ml/孔,随机分为2组(n=18),正常对照组(C组)不作任何处理,吗啡组(M组)加入吗啡,使其终浓度为10μmol/L.于吗啡孵育7 d时应用克隆形成实验测定细胞增殖情况,于吗啡孵育24 h时测定细胞中p53 mRNA、E2F-1 mRNA的表达情况,并采用透射电子显微镜观察细胞超微结构.结果 与C组比较,M组克隆形成率降低,p53 mRNA表达上调,E2F-1 mRNA表达下调(P<0.05).C组细胞核膜完整、核仁及染色质清晰,M组细胞核膜破裂、核仁碎裂,并出现凋亡小体.结论 吗啡可上调人胃癌MGC-803细胞p53基因表达、下调E2F-1基因表达,从而抑制细胞增殖,促进细胞凋亡.
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abstractsObjective To investigate the effect of morphine on the expression of p53 mRNA and E2F-1 mRNA in human gastric carcinoma cell line MGC-803 .Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute, Chinese Academy of Sciences, and cultured in DMEM liquid culture mediun. The cells were seeded in 6-well plates (1 × 103/ml or 2 × 105/ml, 1 ml/well) and divided into 2 groups (n = 18 wells each):group Ⅰ normal control (group C); group Ⅱ was exposed to 10 μmol/L morphine (group M). The proliferation of the cells was determined by colony formation assay at 7 day of incubation with morphine. The expression of p53 mRNA and E2F-1 mRNA was detected and the ulrastructure of the cells examined with transmission electron microscope after being incubated with morphine for 24 h. Results The proliferation of the cells and E2F-1 mRNA expression were significantly lower and p53 mRNA expression was significantly higher in group M than in group C (P < 0.05). The nuclear evelope was intact and the nucleolus and chromosomes were clearly visible in group C, while in group M fragmentation of nuclear envelope and nucleolus and apoptotic bodies were observed. Conclusion Morphine can inhibit the proliferation of the cells and accelerate the cell apoptosis through up-regulating the expression of p53 gene and down-regulating the expression of E2F-1gene in human gastric carcinoma cell line MGC-803.
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