Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, Institute of Developmental Biology, School of Life Science, Shandong University, Ji-nan, China.
Acta Pharmacol Sin. 2013 Jul;34(7):960-8. doi: 10.1038/aps.2013.19. Epub 2013 May 6.
To investigate the effects of 7 novel 1-ferrocenyl-2-(5-phenyl-1H-1,2,4-triazol-3-ylthio) ethanone derivatives on human lung cancer cells in vitro and to determine the mechanisms of action.
A549 human lung cancer cells were examined. Cell viability was analyzed with MTT assay. Cell apoptosis and senescence were examined using Hoechst 33258 and senescence-associated-β-galactosidase (SA-β-gal) staining, respectively. LDH release was measured using a detection kit. Cell cycle was analyzed using a flow cytometer. Intracellular ROS level was measured with the 2',7'-dichlorodihydrofluorescein probe. Phosphorylation of p38 was determined using Western blot.
Compounds 5b, 5d, and 5e (40 and 80 μmol/L) caused significant decrease of A549 cell viability, while other 4 compounds had no effect on the cells. Compounds 5b, 5d, and 5e (80 μmol/L) induced G1-phase arrest (increased the G1 population by 22.6%, 24.23%, and 26.53%, respectively), and markedly increased SA-β-gal-positive cells. However, the compounds did not cause nuclear DNA fragmentation and chromatin condensation in A549 cells. Nor did they affect the release of LDH from the cells. The compounds significantly elevated the intracellular ROS level, decreased the mitochondrial membrane potential, and increased p38 phosphorylation in the cells. In the presence of the antioxidant and free radical scavenger N-acetyl-L-cysteine (10 mmol/L), above effects of compounds 5b, 5d, and 5e were abolished.
The compounds 5b, 5d, and 5e cause neither apoptosis nor necrosis of A549 cells, but exert anti-cancer effect via inducing G1-phase arrest and senescence through ROS/p38 MAP-kinase pathway.
研究 7 种新型 1-二茂铁基-2-(5-苯基-1H-1,2,4-三唑-3-基硫代)乙酮衍生物对体外人肺癌细胞的影响，并确定其作用机制。
检测 A549 人肺癌细胞。MTT 法分析细胞活力。分别用 Hoechst 33258 和衰老相关-β-半乳糖苷酶（SA-β-半乳糖苷）染色检测细胞凋亡和衰老。用检测试剂盒测定乳酸脱氢酶（LDH）释放。用流式细胞仪分析细胞周期。用 2'，7'-二氯二氢荧光素探针测定细胞内 ROS 水平。用 Western blot 测定 p38 的磷酸化。
化合物 5b、5d 和 5e（40 和 80μmol/L）显著降低 A549 细胞活力，而其他 4 种化合物对细胞无影响。化合物 5b、5d 和 5e（80μmol/L）诱导 G1 期阻滞（G1 期细胞分别增加 22.6%、24.23%和 26.53%），并显著增加 SA-β-半乳糖阳性细胞。然而，这些化合物并未导致 A549 细胞的核 DNA 片段化和染色质浓缩。也不影响细胞内 LDH 的释放。这些化合物显著提高了细胞内 ROS 水平，降低了线粒体膜电位，并增加了细胞内 p38 的磷酸化。在抗氧化剂和自由基清除剂 N-乙酰-L-半胱氨酸（10mmol/L）存在的情况下，化合物 5b、5d 和 5e 的上述作用被消除。
化合物 5b、5d 和 5e 既不会引起 A549 细胞凋亡，也不会引起坏死，而是通过 ROS/p38 MAP 激酶通路诱导 G1 期阻滞和衰老发挥抗癌作用。
