苯并呋喃类木脂素衍生物通过抑制细胞周期蛋白质活性诱导MCF-7细胞G2/M期阻滞及凋亡

Induction of G_2/M phase arrest and apoptosis of MCF-7 cells by novel benzofuran lignan via suppressing cell cycle proteins

研究全新合成的苯并呋喃类木脂素化合物2-(3-甲氧基-4-羟基苯基)-4-甲酰基-5-(2-甲氧基羰基乙基)-7-甲氧基苯并[b]呋喃(ERJT-12)的体外抗肿瘤细胞增殖作用及其分子机制。MTT法测定ERJT-12对人肿瘤细胞的抗增殖活性,流式细胞术检测细胞周期分布的改变及细胞凋亡,琼脂糖凝胶电泳检测DNA梯状条带,比色法测定Caspase-3/7和Caspase-6的活性,Western blotting检测细胞周期蛋白质Cdc25c(cell divide cycle 25c)、CDK1(cyclin dependent kinase1)和CyclinB1的改变以及凋亡相关蛋白Bax和Bcl-2的变化。结果显示,ERJT-12对包括多药耐药细胞在内的多种人肿瘤细胞呈现出明显的细胞毒作用。ERJT-12可诱导MCF-7细胞出现明显的G2/M期阻滞及细胞凋亡,细胞中Caspase-3/7和Caspase-6的活性显著升高;CyclinB1蛋白表达下调,Cdc25c和CDK1的活性下降,Bcl-2蛋白被磷酸化。ERJT-12具有显著的抗肿瘤细胞增殖活性,通过抑制细胞周期相关蛋白活性诱导细胞周期阻滞从而触发凋亡,可能成为新的抗肿瘤药物。

In the present study, a newly synthesized benzofuran lignan 4-formyl-2-（4-hydroxy-3-methoxyphenyl）-5-（2-methoxycarbonyethyl）-7-methoxy-benzo[b]furan （ERJT-12） was tested for its antiproliferative activity on human tumor cells. The related mechanisms were also investigated. In vitro growth inhibitory effects of ERJT-12 on various cancer cell lines were determined by MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometry. The integrity of DNA was assessed by agarose gel electrophoresis. Activation of Caspase-3/7 and Caspase-6 was measured by colorimetric assay. The expressions of cell cycle proteins cell divide cycle 25c （Cdc25c）, cyclin dependent kinase 1 （CDK1）, CyclinB1 and apoptosis-related proteins Bax and Bcl-2 were detected by Western blotting. MTT assay showed that ERJT-12 inhibited the proliferation of several cancer cell lines including multidrug resistant cells. MCF-7 cells were markedly arrested at gap2/mitosis （G2/M） phase after treatment with ERJT-12 and progressed into apoptosis. The increased activities of Caspase-3/7 and Caspase-6 in MCF-7 cells were observed. The expression of CyclinB1 was down-regulated. The activities of Cdc25c and CDK1 protein were suppressed and Bcl-2 protein was phosphorylated. ERJT-12 displays potent antiproliferative activity towards cancer cells through suppressing cell cycle proteins, arresting cell cycle at G2/M phase and inducing apoptosis. It might be a novel candidate for cancer therapy.