柴胡皂苷D通过诱导C2-M期阻滞抑制结直肠癌SW480细胞的增殖

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G2-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G2-M周期阻滞(P<0.05)。SSD在mRNA水平下调了G2-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，p21WAF1/CIP1被明显上调。结论:SSD可以通过上调p21WAF1/CIP1的表达诱导G2-M周期阻滞来抑制结直肠肿瘤细胞SW480的增殖。

Saikosaponin -D inhibits the proliferation of colorectal cancer SW480 cell by inducing G2 -Mcell cycle arrest

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 -Mcell 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 -Mcell 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 -Mregulation element p21 WAF1 /CIP1 was up -regulated.Conclusion:SSD inhibits the prolifer-ation of colorectal cancer SW480 cell by up -regulating p21 WAF1 /CIP1 to induce G2 -Mcell cycle arrest.