海洋微生物次级代谢产物1403-B的抗胃癌活性

摘要

目的 筛选能被海洋微生物次级代谢产物1403-B显著抑制的肿瘤细胞,初步探讨其机制,为临床抗肿瘤提供候选药物.方法 用2.5、5.0、10、20、40 μmol/L 1403-B分别处理鼻咽癌细胞株(3种:HONE-1、CNE-2、5-8F),肝癌细胞株(2种:HepG2、Be17402),胃癌细胞株(SGC7901)48 h后,MTT法检测细胞增殖的情况；荧光显微镜观察细胞形态学变化；用Annexin V/PI双染法分析其凋亡情况,Western印迹法检测caspase-3的切割.结果 MTT实验显示,1403-B呈浓度依赖性抑制多种肿瘤细胞的增殖(P＜0.05),其中对胃癌细胞SGC7901的IC5o最低,为(4.12±o.57) μmol/L.1403-B作用SGC7901细胞24h后,出现细胞膜、细胞核皱缩,染色质浓集和形成包裹细胞核片段的凋亡小体等凋亡现象,且呈剂量依赖性增加.Annexin V/PI双染法显示,随着1403-B浓度增加,SGC7901凋亡率升高.1403-B促进SGC7901细胞caspase-3切割.结论 海洋药物1403-B能够通讨诱导SGC7901凋亡而显著抑制其增殖,有望成为一种抗胃癌的新药物.%Objective To confirmed the specific inhibition of 1403-B on the proliferation of tumor cells, to investigate the mechanism for the inhibition. Methods Specific inhibitions of proliferation on different tumor cells such as nasopharyngeal carcinoma cells (HONE-1, CNE-2, 5-8F) , hepatoma cells(HepG2, B7402) and gastric cancer cells( SGC7901) by different concentrations of 1403-B (2. 5, 5. 0, 10, 20, 40 u,mol/L) were detected by MTT assay. Apoptosis induced by 1403-B was measured by fluorescence microscope and Annexin V and propidium iodide ( PI) staining using the Annexin V -FITC Apoptosis Detection Kit. Western blotting assay was used to determine the activation of caspase-3 after 1403-B interacted with SGC7901 cells. Results As detected by MTT assay, ratio of viable SGC7901 cells was significantly declined by 1403-B in a dose-dependent manner and the minimum IC50 was (4. 1 ± 0. 57) jwnol/L in SGC7901. Phenomenons of apoptosis(the membrane and nuclei shrinkage, the chromatin condense and the apoptotic bodies appearance) measured with fluorescence microscope and the percentages of apoptosis tested with flow cytometry in SGC7901 both were increased by 1403-B in a dose-dependent manner. The caspase-3 was activated in the apoptosis of SGC7901 cells. Conclusions 1403-B inhibits the gastric carcinoma cells SGC7901 by induces their apoptosis significantly, suggesting 1403-B will be a potential candidate for treating gastric cancer.