Huang Hong, Zhang Zhen-Xiang, Xu Yong-Jian, Shao Jing-Fang
Department of Respiratory Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, PR China.
Ai Zheng. 2003 Sep;22(9):922-6.
BACKGROUND & OBJECTIVE: Histone deacetylase (HDAC) shows a high expression in many cancer cells and the inhibitor of HDAC1, trichostatin A (TSA), can inhibit the growth of cancer cells. Hypoxia is a common feature of malignant tumors. This paper was designed to investigate the expression of HDAC1 of A549 cell strains in hypoxia condition and the effect of TSA on their proliferation and apoptosis.
The authors designed 1 normoxia group (control group) and 5 hypoxia groups (test groups): hypoxia 6h group (A), TSA + hypoxia 6h (B), hypoxia 12h group (C), hypoxia 24h group (D), TSA + hypoxia 24h (E), hypoxia 48h group (F). The expression of HDAC1 in A549 cells was examined using Western blot analysis. Proliferation, the apoptotic rates of A549 cells and the effect of TSA on them were determined using MTT method, immunohistochemistry, TUNEL method, and flow cytometry. The expression of mRNA of HDAC1 and the effect of TSA on it were determined using reverse transcription-polymerase chain reaction (RT-PCR).
The A values expressed by HDAC1 in A549 cell strains were 138+/-11 in the control group, 78+/-4, 86+/-5, 124+/-3, and 120+/-9 in test groups A, C, D, and F, respectively. The A values of HDAC1mRNA versus the A values of beta-Atin mRNA were 0.68+/-0.03 in the control group, 0.46+/-0.03, 0.45+/-0.02, 0.70+/-0.03, and 0.33+/-0.02 in test groups A, C, D, and F, respectively. The A values of the expression of PCNA in A549 cell strains were 0.13+/-0.03 in the control group, 0.10+/-0.02, 0.11+/-0.02, 0.16+/-0.02, and 0.11+/-0.03 in test groups A, B, D, and E, respectively. The A values of MTT in A549 cell strains were 0.50+/-0.06 in the control group, 0.41+/-0.04, 0.45+/-0.03, 0.59+/-0.02, and 0.45+/-0.03 in test groups A, B, D, and E, respectively. The A values of positive cells of apoptosis in A549 cell strains were 0.16+/-0.04 in the control group, 0.18+/-0.02, 0.18+/-0.05, 0.20+/-0.05, and 0.23+/-0.05 in test groups A, B, D, and E, respectively. The apoptotic rates in A549 cells were 1.11% in the control group, 18.91%,14.30%, 36.99%, and 51.92% in test groups A, B, D, and E, respectively.
The expression of HDAC1 plays an important role in the proliferation and apoptosis of A549 cells, which is regulated by hypoxia. TSA may serve as a new target for therapy of lung cancer.
组蛋白去乙酰化酶（HDAC）在多种癌细胞中高表达，HDAC1抑制剂曲古抑菌素A（TSA）可抑制癌细胞生长。缺氧是恶性肿瘤的常见特征。本文旨在研究缺氧条件下A549细胞株中HDAC1的表达以及TSA对其增殖和凋亡的影响。
作者设计1个常氧组（对照组）和5个缺氧组（试验组）：缺氧6小时组（A）、TSA + 缺氧6小时组（B）、缺氧12小时组（C）、缺氧24小时组（D）、TSA + 缺氧24小时组（E）、缺氧48小时组（F）。采用蛋白质免疫印迹法检测A549细胞中HDAC1的表达。采用MTT法、免疫组织化学法、TUNEL法和流式细胞术检测A549细胞的增殖、凋亡率以及TSA对其的影响。采用逆转录-聚合酶链反应（RT-PCR）检测HDAC1的mRNA表达及TSA对其的影响。
对照组A549细胞株中HDAC1表达的A值为138±11，试验组A、C、D、F中分别为78±4、86±5、124±3、120±9。对照组HDAC1mRNA的A值与β-肌动蛋白mRNA的A值之比为0.68±0.03，试验组A、C、D、F中分别为0.46±0.03、0.45±0.02、0.70±0.03、0.33±0.02。对照组A549细胞株中增殖细胞核抗原（PCNA）表达的A值为0.13±0.03，试验组A、B、D、E中分别为0.10±0.02、0.11±0.02、0.16±0.02、0.11±0.03。对照组A549细胞株中MTT的A值为0.50±0.06，试验组A、B、D、E中分别为0.41±0.04,、0.45±0.03、0.59±0.02、0.45±0.03。对照组A549细胞株中凋亡阳性细胞的A值为0.16±0.04，试验组A、B、D、E中分别为0.18±0.02、0.18±0.05、0.20±0.05、0.23±0.05。对照组A549细胞的凋亡率为1.11%，试验组A、B、D、E中分别为18.91%、14.30%、36.99%和51.92%。
HDAC1的表达在A549细胞增殖和凋亡中起重要作用，且受缺氧调控。TSA可能成为肺癌治疗的新靶点。
