红树林内生真菌来源内酯类化合物D1952抗肿瘤活性研究

Potent antineoplastic effects of a lactone D1952 derived from mangrove microbes

目的探讨红树林老鼠簕内生真菌来源内酯类化合物D1952抗肿瘤作用及分子机制。方法分子对接技术寻找D1952的潜在靶点,四甲基二氮唑蓝(MTT)比色法研究D1952抗肿瘤谱;脱氧核糖核酸末端转移酶介导的缺口末端标记法(TUNEL)、PI染色法、免疫印迹法(Western blotting)检测D1952对细胞周期和凋亡的影响及相关信号通路蛋白的变化。结果细胞周期蛋白激酶2(CDK2)是D1952的潜在靶点;D1952能有效抑制21种肿瘤细胞的增殖,且对MDA-MB-435的半数抑制浓度(IC50)为1.11μmol/L。TUNEL染色可见肿瘤细胞凋亡明显;流式细胞术表明其能将MDA-MB-435细胞阻滞于G1/S期,且D1952能促进PARP的切割、导致p-RB降低和p21的表达增加。结论红树林老鼠簕内生真菌来源化合物D1952具有较好的抗肿瘤活性,并有望成为靶向CDK2的抗肿瘤先导化合物。

Objective To investigate the antineoplastic activity and mechanism of D1952,a lactone derived from mangrove microbes,at both cellular and molecular levels. Methods The target of D1952 was predicted by molecular docking. MTT assay was utilized to study cytotoxicity of D1952 on different tumor cells.Effects of D1952 on cell cycle and apoptosis were tested by terminal deoxynucleotidyl transferase（ TDT）-mediated biotinylated deoxyuridine triphosphate（ d UTP）-biotin nick end labeling（ TUNEL） staining, PI staining and western blotting. Results D1952 effectively inhibited the proliferation of 21 tumor cells with potentially targeting CDK2. The half maximal inhibitory concentration（ IC50） of D1952 in MDA-MB-435 cells was 1. 11 μmol / L. D1952 induced cell apoptosis and apoptosis protein PARP was cleaved leading increased P21 and decreased p-RB. D1952 also induced cell cycle arrest at the G1 / S phase. Conclusion D1952 derived from mangrove microbes displays the antitumor activity, which is expected to be a lead compound targeting CDK2 in relevant tumors.