7-二氟甲氧基-5,4'-二正辛基染料木黄酮通过下调叉头框 M1 抑制胃癌细胞的生长。
7-difluoromethoxyl-5,4'-di-n-octylgenistein inhibits growth of gastric cancer cells through downregulating forkhead box M1.
机构信息
Laboratory of Medical Engineering, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
出版信息
World J Gastroenterol. 2012 Sep 7;18(33):4618-26. doi: 10.3748/wjg.v18.i33.4618.
AIM
To investigate whether the 7-difluoromethoxyl-5, 4'-di-n-octylgenistein (DFOG), a novel synthetic genistein analogue, affects the growth of gastric cancer cells and its mechanisms.
METHODS
A series of genistein analogues were prepared by difluoromethylation and alkylation, and human gastric cancer cell lines AGS and SGC-7901 cultured in vitro were treated with various concentrations of genistein and genistein analogues. The cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cells were incubated by DFOG at different concentrations. The growth inhibitory effects were evaluated using MTT and clonogenic assay. The distribution of the phase in cell cycle was analyzed using flow cytometric analysis with propidium iodide staining. The expression of the transcription factor forkhead box M1 (FOXM1) was analyzed by reverse transcription-polymerase chain reaction and Western blotting. The expression levels of CDK1, Cdc25B, cyclin B and p27(KIP1) protein were detected using Western blotting.
RESULTS
Nine of the genistein analogues had more effective antitumor activity than genistein. Among the tested analogues, DFOG possessed the strongest activity against AGS and SGC-7901 cells in vitro. DFOG significantly inhibited the cell viability and colony formation of AGS and SGC-7901 cells. Moreover, DFOG efficaciously arrested the cell cycle in G2/M phase. DFOG decreased the expression of FOXM1 and its downstream genes, such as CDK1, Cdc25B, cyclin B, and increased p27(KIP1) at protein levels. Knockdown of FOXM1 by small interfering RNA before DFOG treatment resulted in enhanced cell growth inhibition in AGS cells. Up-regulation of FOXM1 by cDNA transfection attenuated DFOG-induced cell growth inhibition in AGS cells.
CONCLUSION
DFOG inhibits the growth of human gastric cancer cells by down-regulating the FOXM1 expression.
目的
研究新型大豆异黄酮类似物 7-二氟甲氧基-5,4'-二正辛基染料木黄酮（DFOG）是否影响胃癌细胞的生长及其机制。
方法
通过二氟甲基化和烷基化合成了一系列大豆异黄酮类似物，并用不同浓度的染料木黄酮及其类似物处理体外培养的人胃癌细胞系 AGS 和 SGC-7901。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法测定细胞活力。用不同浓度的 DFOG 孵育细胞，用 MTT 和集落形成实验评估生长抑制作用。用碘化丙啶染色的流式细胞术分析细胞周期时相分布。用逆转录-聚合酶链反应和 Western blot 分析叉头框 M1（FOXM1）转录因子的表达。用 Western blot 检测 CDK1、Cdc25B、cyclin B 和 p27(KIP1) 蛋白的表达水平。
结果
9 种大豆异黄酮类似物的抗肿瘤活性强于染料木黄酮。在测试的类似物中，DFOG 对 AGS 和 SGC-7901 细胞的体外活性最强。DFOG 显著抑制 AGS 和 SGC-7901 细胞的细胞活力和集落形成。此外，DFOG 有效地将细胞周期阻滞在 G2/M 期。DFOG 降低了 FOXM1 及其下游基因如 CDK1、Cdc25B、cyclin B 的表达，并增加了 p27(KIP1) 的蛋白水平。DFOG 处理前用小干扰 RNA 敲低 FOXM1 导致 AGS 细胞的生长抑制增强。用 cDNA 转染上调 FOXM1 减弱了 DFOG 诱导的 AGS 细胞生长抑制。
结论
DFOG 通过下调 FOXM1 表达抑制人胃癌细胞的生长。
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