Effects of the lipidperoxidation product 4-hydroxynonenal and related aldehydes on proliferation and viability of cultured Ehrlich ascites tumor cells.
Effects of the lipidperoxidation product 4-hydroxynonenal and related aldehydes on proliferation and viability of cultured Ehrlich ascites tumor cells.
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DOI：
10.1016/0006-2952(85)90428-9
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发表时间：
1985-11
影响因子：
5.8
通讯作者：
S. Hauptlorenz;H. Esterbauer;W. Moll;R. Pümpel;E. Schauenstein;B. Puschendorf
中科院分区：
文献类型：
--
作者：
S. Hauptlorenz;H. Esterbauer;W. Moll;R. Pümpel;E. Schauenstein;B. Puschendorf
脂质过氧化产物4-羟基壬烯醛和其他几种同源非生物源性醛抑制培养的埃利希腹水肿瘤细胞增殖的机制已被研究。细胞（5 × 10−4/ml）在添加10或20 μM -羟基壬烯醛的最低基本培养基中孵育，可将36小时的细胞计数降低至65和控制值的30%。生长速度的降低很可能是由于DNA合成受阻。在暴露于4-羟基壬烯醛（20 μM， 8小时）之前，用[3H]-胸腺嘧啶脉冲标记的细胞显示DNA的特定放射性没有变化，这表明在醛的存在下发生了淋巴结新合成。在没有乙醛的情况下，DNA的放射性降低了25%。在10 μM或20 μM的4-羟基壬烯醛存在下孵育2小时，使[3H]-胸腺嘧啶在hclo4不溶性部分的掺入率降低到对照组的85%和50%，但对[3H]-胸腺嘧啶和86rb的摄取没有影响。此外，通过台锥蓝排斥技术检测细胞培养结果显示，20 μM - 4-羟基不引起细胞死亡。4-羟基壬烯醛对巯基的高反应性表明，醛通过与DNA聚合酶的SH基团相互作用抑制DNA合成。当甲醛浓度为50 μM时，对DNA合成的特异性作用消失，导致30%（6小时）和95%（36小时）的细胞死亡。4-羟基壬烯醛的细胞抑制指数（CI）为3.0，即50%台盼蓝阳性细胞的浓度/ 50%抑制细胞生长的浓度。其他链长为5、6、7、8、10和11个碳原子的同源4-羟基烯醛也能抑制细胞生长。CI在1.20 ~ 1.94之间变化，表明非生物源性4-羟基烯醛对增殖的特异性作用明显低于生物源性4-羟基壬烯醛。4-羟基壬烯醛与谷胱甘肽和半胱氨酸的Michael加合物毒性比游离醛低近一个数量级，但CI（2.41半胱氨酸加合物，2.06谷胱甘肽加合物）并没有提高，因为生长抑制作用也降低了。
The mechanism by which the lipid peroxidation product 4-hydroxynonenal and several other homologous, yet non biogenic aldehydes inhibit proliferation of cultured Ehrlich ascites tumor cells has been studied. Incubation of cells (5 × 10−4/ml) in a minimum essential medium supplemented with 10 or 20 μM 4-hydroxynonenal reduces the 36-hr cell count to 65 and 30% of the control value. The reduced growth rate is most likely due to a blockage of the DNA synthesis. Cells labelled by a [3H]-thymidine pulse prior to exposure to 4-hydroxynonenal (20 μM, 8 hr) showed no change of the specific radioactivity of the DNA, indicating that node novosynthesis occured in the presence of the aldehyde. In the absence of the aldehyde the specific radioactivity of the DNA decreased by 25%. A 2-hr incubation in the presence of 10 or 20 μM of 4-hydroxynonenal reduced [3H]-thymidine incorporation into the HClO4insoluble fraction to 85 and 50% of the controls, but had no effect of the [3H]-thymidine and86Rb uptake. Moreover, examination of the cell cultures by the Trypan Blue exclusion technique revealed that 20 μM 4-hydroxynonal does not cause cell death. The high reactivity of 4-hydroxynonenal towards sulfhydryl groups suggests that the aldehyde inhibits DNA synthesis by interacting with a functional SH group of DNA polymerase. The specific action on DNA synthesis is abolished at an aldehyde concentration of 50 μM, which leads to 30% (6 hr exposure) and 95% (36 hr exposure) of dead cells.The cytostatic index (CI), i.e. concentration at 50% Trypan Blue positive cells/concentration at 50% inhibition of cell growth deducted from the dose effect curves is 3.0 for 4-hydroxynonenal. The other homologous 4-hydroxyalkenals with chain length of 5, 6, 7, 8, 10 and 11 carbon atoms also inhibit cell growth. The CI varied from 1.20 to 1.94, indicating that these non biogenic 4-hydroxyalkenals have a distinctively lower specific effect on proliferation than the biogenic 4-hydroxynonenal.The Michael adducts of 4-hydroxynonenal with glutathione and cysteine were nearly one order of magnitude less toxic than the free aldehyde, the CI (2.41 cysteine adduct, 2.06 glutathione adduct), however, were not improved since the growth inhibitory action was also reduced.