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目的观察人胚胎肺成纤维细胞(HLF)正常及缺陷细胞在低剂量氢醌(hydroquinone,HQ)处理后能否诱导适应性反应,探讨PARP1蛋白缺陷与正常细胞诱导适应性反应与微核形成及细胞周期变化的关系,进一步阐明适应性反应形成机制。方法用低剂量HQ预处理细胞后,观察细胞和DNA对大剂量HQ攻击的适应性反应,从微核率及核异常率、细胞周期等方面测定细胞变化情况。结果在细胞整体活力水平,0.001～0.050μmolLHQ预处理可以提高细胞对攻击剂量80.0μmolL的耐受性;HLF、转染绿色荧光蛋白真核表达载体的HLF(HLFC)和转染hPARP1基因反义RNA真核表达载体的HLF(HLFP)细胞经HQ低剂量预处理及大剂量攻击后,各剂量组不同程度出现含微核和核异常细胞,G1、G2、S期细胞数分布均不相同。与同种细胞仅大剂量攻击比较,0～0.050μmolLHQ预处理的HLF、HLFC及HLFP细胞微核率及核异常率差异均有统计学意义(P<0.05)。HQ预处理的HLF和HLFC细胞均出现G2期阻滞;HLFP细胞表现为G2期阻滞,1.000、2.000μmolL出现G1期阻滞。结论在低剂量HQ作用下,hPARP1蛋白缺陷细胞和正常细胞一样有适应性反应,但低于正常细胞,说明hPARP1蛋白在细胞适应性反应方面起重要作用,适应性反应与细胞周期调控等有关。
Objective To observe whether adaptive response induced by low-dose hydroquinone (HQ) can be induced in normal and defective cells of human embryonic lung fibroblasts (HLF), and to investigate the relationship between PARP1 deficiency and normal cells-induced adaptive responses and micronuclei formation Cell cycle changes in the relationship between further clarify the formation of adaptive response mechanism. Methods After pretreatment of cells with low dose of HQ, the adaptive response of cells and DNA to high dose HQ challenge was observed. The changes of cells were measured from micronucleus rate, nuclear anomalous rate and cell cycle. RESULTS: Pretreatment with 0.001 ~ 0.050μmolLHQ could increase the tolerance of cells to challenge dose of 80.0μmolL. HLF, HLF transfected with green fluorescent protein eukaryotic expression vector and antisense RNA transfected with hPARP1 gene The HLF (HLFP) cells of eukaryotic expression vector were pretreated with low dose of HQ and challenged with high dose. The micronuclei and abnormal nuclei of cells in different dose groups appeared in different dose groups. The cell numbers in G1, G2 and S phase were all different. Compared with high-dose group, the micronucleus rate and nuclear anomalous rate of HLF, HLFC and HLFP cells pretreated with 0 ~ 0.050μmolLHQ were all significantly different (P <0.05). G2 phase arrest occurred in both HLF and HLFC cells pretreated with HQ; G2 phase arrest was observed in HLFP cells, and G1 arrest occurred at 1.000 and 2.000 μmolL. Conclusion Under the action of low dose of HQ, hPARP1-deficient cells have the same adaptive response as normal cells but lower than normal cells, indicating that hPARP1 protein plays an important role in cell adaptive response. Adaptive response is related to cell cycle regulation and so on.