【作者】 王国红；

【导师】 李祺福；

【作者基本信息】 厦门大学 ， 细胞生物学， 2005， 硕士

【摘要】 本研究应用课题组所建立的人成骨肉瘤MG-63细胞终末分化模型,系统研究中药有效成分人参皂甙Rg1、肉桂酸、丹参酮ⅡA以及人参皂甙Rg1+肉桂酸、人参皂甙Rg1+肉桂酸+丹参酮ⅡA组合对人成骨肉瘤MG-63细胞终末分化的诱导作用,以进一步鉴定中药有效成分对肿瘤细胞终末分化的诱导效果并初步探讨其诱导肿瘤细胞终末分化的作用机理。实验结果显示,经人参皂甙Rg1、肉桂酸、丹参酮ⅡA及其组合处理后,MG-63细胞增殖受到不同程度的抑制,抑制率分别为22.06%、54.94%、52.10%、59.23%和72.37%。细胞倍增时间由对照组的47.96h分别延长至52.69h、67.29h、65.27h、70.88h和89.39h。细胞周期检测结果显示,经人参皂甙Rg1、肉桂酸、丹参酮ⅡA及其组合处理后,细胞均发生G0/G1期阻滞,G0/G1期细胞比例由对照组的47.5%分别上升到56.6%、59.8%、59.5%、67.9%和72.5%,S期细胞比例则由对照组的20.0%分别下降至11.8%、10.3%、9.0%、8.2%和7.3%。光镜和透射电子显微镜的观察结果显示,经人参皂甙Rg1、肉桂酸、丹参酮ⅡA及其组合处理后的MG-63细胞出现了细胞形态规则,大小趋于一致,细胞体积增大,核质比例减小,细胞核形状规则、核内异染色质减少、常染色质增多,线粒体数量增多、形态较为一致,粗糙内质网增多、游离核糖体增多而多聚核糖体减少,细胞表面微绒毛减少等变化。成骨肉瘤细胞终末分化标志物Ⅰ型胶原、骨钙蛋白、骨粘素表达增强、钙沉积增加,有典型骨结节形成且数目增多,其中联合处理组的效果强于单独处理组。免疫细胞化学检测结果显示,经人参皂甙Rg1、肉桂酸、丹参酮ⅡA及其组合处理之后,细胞增殖分化调控相关的癌基因c-fos和c-myc表达活性降低,抑癌基因Rb和p27的表达活性升高。研究结果表明,中药有效成分人参皂甙Rg1、肉桂酸、丹参酮ⅡA及其组合能够有效抑制人成骨肉瘤MG-63细胞的增殖活动,将细胞周期阻滞更多还原

【Abstract】 In this study, we used the terminal differentiation model of human osteosarcoma MG-63 cells built by our research group to investigate the induction effects of ginsenoside Rg1, cinnamic acid , tanshinoneⅡA and ginsenoside Rg1 + cinnamic acid, ginsenoside Rg1 + cinnamic acid + tanshinoneⅡA on the terminal differentiation of human osteosarcoma MG-63 cells. The aim is to further identify the induction effects of effective ingredients of Chinese traditional medicine on terminal differentiation of tumour cells and explore the machamism of terminal differentiation induction primarily.Our results revealed that the proliferation of MG-63 cells was inhibited differently after the treatments of ginsenoside Rg1, cinnamic acid, tanshinoneⅡA and their combinations. The inhibitory rates were 22.06%、54.94%、52.10%、59.23% and 72.37% and the doubling time was prolonged from 47.96h to 52.69h、67.29h、65.27h、70.88h and 89.39h. Flow cytometry analysis showed that MG-63 cells were arrested in G0/G1 phase after the treatments. The proportion of cells in G0/G1 phase rose from 47.5% to 56.6%、59.8%、59.5%、67.9% and 72.5% while the proportion of cells in S phase declined from 20.0% to 11.8%、10.3%、9.0%、8.2% and 7.3%.With the light microscope and transmission electron microscope, we observed that the MG-63 cells had undergone malignant morphology and ultrastructure reversion after the treatments. The morphology of the cells was regular and the size of them tended to be similar. The size of the cells increased. And the proportion of nucleus to cytoplasm decreased. The morphology of the nucleus was regular. The heterochromatin decreased while the euchromatin increased. The number of mitochondrions increased and their size appeared similar. Rough endoplasmic reticulum and free ribosomes increased while polyribosomes decreased. The更多还原