β-榄香烯体内外对人肺癌细胞株的作用及其机制

Effect and Its Mechanism of β-elemene on Human Lung Cancer SPC-A-1 Cell Lines in Vivo and in Vitro

目的：系统研究β-榄香烯体内外抗肺癌药理学作用及其作用机制。方法：体外抗肿瘤实验采用MTT法,检测浓度12.5~200μmol·L-1的β-榄香烯对人肺癌细胞株SPC-A-1,A549及人正常胚胎肺成纤维细胞MRC-5的细胞增殖抑制率;抗肿瘤机制研究采用Hoechst 33258染色,观察细胞凋亡时细胞核的形态学变化,流式细胞术FITC-Annexin V/碘化丙啶（PI）双标记染色法检测细胞凋亡率。体内以裸鼠移植瘤为模型,肿瘤生长抑制率为指标,检测β-榄香烯对荷瘤裸鼠肿瘤生长的抑制作用。结果：β-榄香烯作用48 h时SPC-A-1,A549的50%抑制浓度（IC50）分别为126.0,134.1μmol·L-1,但对人胚胎肺成纤维细胞MRC-5有促进增殖作用,表明β-榄香烯对肺癌细胞具有选择性抑制作用。β-榄香烯对人肺癌细胞株SPC-A-1增殖的抑制作用呈时间和浓度依赖性,干预24,48,72 h的IC50值分别为549.7,126.0,21.2μmol·L^-1;Hoechst 33258染色后倒置荧光显微镜下观察可见β-榄香烯孵育后出现凋亡细胞核固缩,浓染,着色深而呈亮蓝色;流式细胞术检测结果显示,β-榄香烯可诱导SPC-A-1细胞出现不同程度的凋亡。给药剂量为45,135 mg·kg^-1的β-榄香烯在体内对裸鼠移植性人肺癌SPC-A-1细胞的增殖抑制率分别为34.01%,49.57%。结论：β-榄香烯具有选择性抗肺癌作用,提示其在发挥抗肿瘤作用的同时其毒副作用小,而且该作用可能机制是通过诱导肺癌细胞凋亡实现的。

Method： The in vitro Objective ： To study effects of beta-elemene the effect and mechanism （ 12.5-200 μmol·L^ -1 ） on A549 ceils and human diploid fibroblasts MRC-5 cell prolifera n of beta-elemene in human lung cancer vivo and in vitro. SPC-A-1 cell line, were observed by MTT method; the morphological changes of apoptotic cells were detected by Hoechst 33258; the cells stained by Annexin V and propidium iodide （PI） were observed by flow cytometry for apoptosis, cell morphological changes were observed electron microscope. Its in vivo anti-tumor effect on transplanted human SPC-A-1 tumor cells in nude mice was tected by conventional methods. Result： Fifty percent inhibition concentration （ICs0） of the above-mentioned 2kinds of lung cancer cells by beta-elemene was 126.0, 134.1 μmol·L^-1, but the effect on human diploid fibroblasts MRC-5 cells was promoting growth. The inhibition effects of beta-elemene on human lung cancer cell line SPC-A-1 depended on the duration and concentrations. IC50 at the 24, 48, 72 h was 549.7, 126.0, 21.2 μmol·L^-1, respectively. Apoptotic body in apoptotic cells could be observed by Hoechst 33258 dying. Flow cytometry showed that beta-elemene could induce SPC-A-1 cell apoptosis distinctly at different levels. The growth inhibition rates of beta-elemene （45, 135 mg ·kg ^-1 ） on transplanted human lung cancer cell line SPC-A-1 ceils in vivo were 34.01% , 49.57%. Conclusion： Beta-elemene has an antitumor effect between 12.5-200 μmol·L^-1. The mechanism of anti-tumor effect is possibly achieved by inducing apoptosis of tumor cells.