Miki H, Kasprzak K S, Kenney S, Heine U I
Laboratory of Comparative Carcinogenesis, National Cancer Institute, Frederick, MD 21701-1013.
Carcinogenesis. 1987 Nov;8(11):1757-60. doi: 10.1093/carcin/8.11.1757.
The level of gap-junctional (cell-cell) communication was studied by the radioisotope transfer technique in NIH 3T3 cells exposed to NiSO4, MgSO4, or both salts combined. Monolayered NIH 3T3 donor cells were labeled with [3H]-uridine for 3 h and then co-cultured with non-labeled recipient NIH 3T3 cells for 3 h in the presence of 0.5-20 mM NiSO4, 1.0-100 mM MgSO4, or 5 mM NiSO4 plus 1.0-100 mM MgSO4. 12-O-tetradecanoylphorbol-13-acetate (TPA), 16-160 pM, served as a positive control. The exposed cells were fixed with 2.5% glutaraldehyde and processed for autoradiography. The cell-cell communication rate was based on the number of radioactive recipient cells in relation to the total number of recipient cells for 100 donor cells. NiSO4 disrupted cell-cell communication in a dose-related manner from 98% of the base value at 0.5 mM NiSO4 to 2% at 5 mM NiSO4. Cell viability was not affected by 0.5-5 mM NiSO4. The inhibitory action of 5 mM NiSO4 could be partially prevented by 5.0-100 mM MgSO4. However, MgSO4 did not prevent the inhibition by TPA. The results indicate that NiSO4 is capable of inhibiting cell-cell communication at concentrations that do not cause cytotoxic effects in NIH 3T3 cells during a 3-h period. In this respect NiSO4 resembles such classical tumor promoters like TPA. The antagonism by magnesium of the nickel-induced inhibition of cell-cell communication may indicate a contributory mechanism by which magnesium counteracts the carcinogenicity of nickel in vivo.
采用放射性同位素转移技术,对暴露于硫酸镍、硫酸镁或两种盐组合的NIH 3T3细胞中的间隙连接(细胞-细胞)通讯水平进行了研究。将单层NIH 3T3供体细胞用[³H]-尿苷标记3小时,然后在0.5 - 20 mM硫酸镍、1.0 - 100 mM硫酸镁或5 mM硫酸镍加1.0 - 100 mM硫酸镁存在的情况下,与未标记的受体NIH 3T3细胞共培养3小时。16 - 160 pM的12 - O - 十四酰佛波醇 - 13 - 乙酸酯(TPA)用作阳性对照。将暴露的细胞用2.5%戊二醛固定并进行放射自显影处理。细胞-细胞通讯率基于相对于100个供体细胞的受体细胞总数的放射性受体细胞数量。硫酸镍以剂量相关的方式破坏细胞-细胞通讯,从0.5 mM硫酸镍时为基础值的98%降至5 mM硫酸镍时的2%。0.5 - 5 mM硫酸镍不影响细胞活力。5.0 - 100 mM硫酸镁可部分阻止5 mM硫酸镍的抑制作用。然而,硫酸镁不能阻止TPA的抑制作用。结果表明,硫酸镍在3小时内能够在不引起NIH 3T3细胞细胞毒性作用的浓度下抑制细胞-细胞通讯。在这方面,硫酸镍类似于TPA等经典肿瘤促进剂。镁对镍诱导的细胞-细胞通讯抑制的拮抗作用可能表明镁在体内抵消镍致癌性的一种促成机制。