Effects of the histone deacetylase inhibitor trichostatin A on nuclear texture and c-jun gene expression in drug-sensitive and drug-resistant human H69 lung carcinoma cells
Effects of the histone deacetylase inhibitor trichostatin A on nuclear texture and c-jun gene expression in drug-sensitive and drug-resistant human H69 lung carcinoma cells
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DOI：
10.1002/cyto.a.20088
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发表时间：
2004-12-01
期刊：
影响因子：
3.7
通讯作者：
Dufer, J
中科院分区：
文献类型：
--
作者：
El-Khoury, V;Gomez, D;Dufer, J
背景：通过图像细胞术对染色质模式进行纹理分析可用于癌症诊断和预后的开发和完善以及治疗的后续治疗。然而，人们对这些模式背后的生物学机制知之甚少。表观遗传机制作为历史性翻译后修饰，尤其是组蛋白乙酰化，可能在这些染色质模式的确定中发挥重要作用，进而影响核纹理测量。 方法：本研究探讨了组蛋白脱乙酰酶抑制剂曲古抑菌素 A (TSA) 治疗对化疗敏感和耐药的人类细胞系核纹理的影响。将小细胞肺癌 H69 细胞及其对依托泊苷具有抗性的变体 H69-VP 与 100 ng/ml TSA 一起孵育 0 至 24 小时。通过图像细胞术评估核纹理，并与通过蛋白质印迹法测量的组蛋白 H4 乙酰化水平以及通过逆转录和实时聚合酶链反应评估 c-jun 基因的表达进行比较。 结果：TSA 处理诱导两种细胞系中组蛋白 H4 乙酰化水平的增加。然而，在染色质纹理水平上，敏感的 H69 细胞在长达 24 小时内表现出渐进的染色质解浓缩，而耐药的 H69-VP 显示出快速（8 小时）但短暂的变化。同样，在 TSA 处理的 H69 细胞中，c-jun 的表达有规律地增加。在 H69-VP 细胞中，在治疗 12 小时内也观察到增加，随后在治疗 24 小时后减少。结论：核纹理分析似乎是检测组蛋白脱乙酰酶抑制剂 TSA 在 H69 细胞系中诱导的染色质模式改变的敏感技术，并能够观察在此期间化疗药物敏感细胞和耐药细胞之间的染色质模式差异。 治疗。当c-jun基因表达被分析为对表观遗传控制敏感的基因时，这些结构差异似乎与基因表达相关。 (C) 2004 Wiley-Liss, Inc.
Background: Texture analysis of chromatin patterns by image cytometry can be used in the development and refinement of diagnosis and prognosis of cancers and in the follow-up of therapies. However, little is known about the biological mechanisms underlying these patterns. Epigenetic mechanisms as historic posttranslational modifications and particularly histone acetylation could play a major role in the determination of these chromatin patterns and then influence nuclear texture measurements.Methods: This study examined the consequences of treatment by the histone deacetylase inhibitor trichostatin A (TSA) on the nuclear texture in human cell lines sensitive and resistant to chemotherapy. Small cell lung carcinoma H69 cells and their variant H69-VP, which is resistant to etoposide, were incubated with 100 ng/ml of TSA for 0 to 24 h. Nuclear texture was evaluated by image cytometry and compared with the histone H4 acetylation level measured by western blotting and expression of c-jun gene evaluated by reverse transcription and real-time polymerase chain reaction.Results: TSA treatment induced an increase in histone H4 acetylation level in both cell lines. However, at the level of chromatin texture, sensitive H69 cells displayed a progressive chromatin decondensation up to 24 h, whereas resistant H69-VP showed rapid (8 h) but transient changes. Similarly, expression of c-jun increased regularly in TSA-treated H69 cells. In H69-VP cells, an increase was also observed up to 12 h followed by a decrease after 24 h of treatment.Conclusions: Analysis of nuclear texture appeared to be a sensitive technique to detect chromatin pattern alterations induced by the histone deacetylase inhibitor TSA in the H69 cell line and enabled the observation of chromatin pattern discrepancies between chemotherapeutic drug-sensitive and drug-resistant cells during this treatment. When c-jun gene expression was analyzed as gene sensitive to epigenetic control, these textural differences seemed to be correlated to gene expression. (C) 2004 Wiley-Liss, Inc.