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背景:低剂量活性氧可以促进正常细胞和肿瘤细胞增殖、分化,但目前对此过程中所涉及的信号途径及其与细胞敏感时相的关系尚认识不足。目的:通过阻断细胞内NF-κB途径和诱导不同细胞时相,观察过氧化氢促进HepG2细胞增殖与NF-κB途径和细胞敏感时相的关系,为抑制、干预肿瘤细胞的生长及改善患者预后功能提供理论依据。设计:完全随机实验研究。单位:解放军第四军医大学预防医学系毒理学教研室。对象:实验于2004-04/2004-08在解放军第四军医大学预防医学系毒理学教研室完成,对象为人肝癌细胞株HepG2细胞,由解放军第四军医大学基础部病理学教研室惠赠。方法:以不同剂量过氧化氢直接作用于培养的HepG2细胞,筛选能够促进HepG2细胞增殖的过氧化氢浓度。用50μmol/L的过氧化氢分别处理经NF-κB抑制剂(PDTC)预处理的HepG2细胞和用胸腺嘧啶核苷同步化得到的G1,G2/M,S期HepG2细胞。主要观察指标:细胞分别经过氧化氢处理1,24,48h后,通过四甲基偶氮唑蓝(MTT)比色法检测各组细胞吸光度值,比较细胞增殖活性。结果:50μmol/L的过氧化氢作用HepG2细胞24h后,MTT实验所测吸光度值与对照组相比显著升高(t=0.001,P<0.01)。20μmol/LPDTC预处理HepG2细胞2h后,给予50μmol/L过氧化氢作用细胞,MTT实验所测吸光度值与对照组相比差异
BACKGROUND: Low dose reactive oxygen species (ROS) can promote the proliferation and differentiation of normal cells and tumor cells. However, the current signaling pathways involved in this process and their relationship to the sensitive phase of cells are not well understood. OBJECTIVE: To observe the effect of hydrogen peroxide on promoting the proliferation of HepG2 cells and the relationship between NF-κB pathway and cell-sensitive phase by blocking NF-κB pathway and inducing different cell phases, in order to inhibit the growth and improve the growth of tumor cells Prognostic function to provide a theoretical basis. Design: Completely randomized experimental study. Unit: Department of Toxicology, Department of Preventive Medicine, Fourth Military Medical University. PARTICIPANTS: The experiment was performed at Department of Toxicology, Department of Preventive Medicine, Fourth Military Medical University of Chinese PLA from April 2004 to August 2004. The target was HepG2 human hepatoma cell line, which was kindly donated by Department of Pathology, Fourth Military Medical University of PLA. Methods: HepG2 cells were cultured with different doses of hydrogen peroxide directly and the concentration of hydrogen peroxide that could promote the proliferation of HepG2 cells was screened. HepG2 cells pretreated with NF-κB inhibitor (PDTC) and G1, G2 / M, S phase HepG2 cells synchronized with thymidine were treated with 50 μmol / L hydrogen peroxide. MAIN OUTCOME MEASURES: Cells were treated with hydrogen peroxide for 1, 24 and 48 hours, respectively, and the cell absorbance values were determined by MTT assay to compare the cell proliferative activity. Results: After treated with 50μmol / L H2O2 for 24h, the absorbance value of MTT assay was significantly higher than that of the control group (t = 0.001, P <0.01). HepG2 cells were pretreated with 20μmol / LPDTC for 2h and then treated with 50μmol / L H2O2. The difference between the absorbance values measured by MTT assay and the control group