柴胡皂苷D通过诱导C_2-M期阻滞抑制结直肠癌SW480细胞的增殖  

Saikosaponin- D inhibits the proliferation of colorectal cancer SW480 cell by inducing G_2-M cell cycle arrest

作　　者：杨坤荣[1] 王健生[1] 

机构地区：[1]西安交通大学第一附属医院肿瘤外科,陕西西安710061

出　　处：《现代肿瘤医学》2016年第9期1353-1357,共5页Journal of Modern Oncology

摘　　要：目的:观察柴胡皂苷D(SSD)对于结直肠癌肿瘤细胞增殖的影响并初步探索其潜在的分子机制。方法:运用MTT、细胞计数试验及克隆形成试验观察SSD对细胞增殖的影响。流式细胞仪检测SSD对细胞凋亡及周期分布情况的影响。RT-PCR和Western-blot技术检测G_2-M周期阻滞关键调控因子在SW480细胞中的表达。结果:MTT试验发现SSD能够显著抑制结直肠癌肿瘤细胞SW480的增殖,而对正常结直肠细胞FHC的增殖无明显影响。细胞计数试验和克隆形成试验进一步验证了SSD对于SW480细胞增殖的影响(P<0.05)。流式细胞术检测发现SSD不影响细胞的凋亡(P>0.05),但却显著诱导G_2-M周期阻滞(P<0.05)。SSD在mRNA水平下调了G_2-M周期调控因子CCNA1、CCNA2、CCNB1和CCNB2的表达(P<0.05);在蛋白水平,CCNA2、CCNB1和p34/cdc2明显被下调,p-H3S10上调,p21^(WAF1/CIP1)被明显上调。结论:SSD可以通过上调p21^(WAF1/CIP1)的表达诱导G_2-M周期阻滞来抑制结直肠肿瘤细胞SW480的增殖。Objective： To observe the anti - proliferation capacity of saikosaponin - D （SSD） on colorectal cancer cell and normal colorectal cell and further explore the underlying mechanisms. Methods：Using MTT, cell counting and clone formation assay to assess the anti - growth effects of SSD. Flow - cytometer was used to check apoptosis and cell cycle. RT - PCR and Western - blot were used to check the pivotal elements in the regulation of G2 - M cell cycle arrest. Results ： SSD significantly inhibited the proliferation of SW480 rather than FHC （ P 〈 0.05 ）. It had no impact on cell apoptosis （P 〉 0.05 ） , but significantly induced G2 -M cell cycle arrest as determined by Flow- cytomete （P 〈 0.05）. SSD down - regulated the expression of CCNA1, CCNA2, CCNB1 and CCNB2 in mRNA level by RT - PCR （P 〈 0.05）. In protein level, CCNA2, CCNB1 and p34/cdc2 were down - regulated, p - H3S10 was up - regulated, while another pivotal G2 - M regulation element p21 WAF4/CIP1 was up - regulated. Conclusion：SSD inhibits the proliferation of colorectal cancer SW480 cell by up - regulating p21WAF4/CIP1 to induce G2 - M cell cycle arrest.

关 键 词：结直肠癌 柴胡皂苷D G2-M周期阻滞 细胞增殖 

分 类 号：R735.3[医药卫生—肿瘤]