摘要
目的观察蟾毒灵抗高转移潜能肝癌细胞增殖凋亡作用。方法体外培养人高转移潜能HCC—LM3肝癌细胞，分别加入不同浓度蟾毒灵（0．02、0．04、0．08mg／L）和5-氟尿嘧啶（5-Fu，1．05、12．34、25．79mg／L）作用肝癌细胞24、48、72h后，采用细胞计数试剂盒（CCK-8）、流式细胞技术测定肝癌细胞生长、增殖周期变化和细胞凋亡。结果同-时问点随蟾毒灵浓度增加对肝癌细胞生长抑制率逐渐增加（F=14．823、15．623、4．737，P〈0．05），同-浓度随作用时间延长蟾毒灵对肝癌细胞生长抑制率逐渐增加（F=44．757、43．256、19．682、17．112、50．765、31．214，P〈0．05）。与对照组相比，蟾毒灵浓度增加导致G．期肝癌细胞比例逐渐降低[IC50组：（43．96±3．52）％比（70．19±4．70）％，IC50组：（40．99±5．72）％比（70．19±4．70）％，F=23．924，P〈0．05]，G2＋s期肝癌细胞比例逐渐增加[IC，。组：（56．04±3．52）％比（29．81±4．70）％，IC70组：（59．01±5．72）％比（29．81±4．70）％，F=23．924，P〈0．05]。不同作用时间随蟾毒灵浓度增加，肝癌细胞凋亡率逐渐增加（F=6．817、46．639、77．658，P〈0．05），且作用48h[Ic50组：（34．01±4．62）％比（22．02±1．29）％，IC50组：（47．60±4．41）％比（3I．90±4．95）％，t=-6．381，P〈0．05]和72h蟾毒灵诱导肝癌细胞凋亡率高于5-FU[IC50组：（52．01±6．63）％比（32．65±3．69）％，IC70组：（74．31±6．18）％比（47．85±4．52）％，t=5．729，P〈0．05]。结论蟾毒灵通过阻滞肝癌细胞增殖于s期和G：期诱导细胞凋亡。
Objective To investigate the inhibitory effects of Bufalin on human hepatocellular car- cinoma cells with high metastatic potential. Methods The human hepatoma cells with high metastatic po- tential （ HCC - LM3 ） were cultured in vitro. The bufalin was respectively added into culture fluid with va- rious concentration of 0. 02, 0. 04, 0. 08 mg/L. The 5 -fluorouracil （5 -Fu） was respectively added into culture fluid with various concentration of 1.05, 12. 34, 25.79 mg/L. The cell growth, proliferative cycle and apoptosis were respectively determined by eel1 counting kit -8 （CCK -8 ） assay and flow eytometry technique at 24, 48, 72 h. Results The growth inhibitory rate of HCC - LM3 cells gradually increased with the increase of bufalin concentration at the same time （ F = 14. 823, 15. 623, 4. 737, P 〈 0. 05 ） or with the time extension of bufalin at the same concentration （F= 44. 757, 43. 256, 19. 682, 17. 112, 50. 765, 31. 214, P 〈0. 05）. Compared with control group, the cell percentage of HCC - LM3 cells at Gl phase gradually decreased with the increase of bufalin concentration [ IC50 ： （43.96 ± 3.52 ） % vs. （70. 19 ± 4.70） % , IC70 ： （40. 99 ± 5.72） % vs. （70. 19 ±4.70） % , F = 23. 924, P 〈 0. 05 ] , and the cell percentage of HCC - LM3 cells at G2 plus S phases gradually increased with the increase of bufalin concentration [IC50：（56.04±3.52）% vs. （29,81 ±4.70）%,IC70：（59.01 ±5.72）% vs. （29.81 ＋4.70）%,F= 23. 924, P 〈 0. 05 ]. The apoptotic rate of HCC - LM3 cells gradually increased with the increase of bufalin concentration at various time （F = 6. 817, 46. 639, 77. 658, P 〈 0. 05）. The apoptotic rate of HCC -LM3 cells induced by bufalin was higher than that of 5 - Fu at 48 h and 72 h [ IC50 ： （ 34. 01±4. 62 ） % vs. （22.02±1.29）%,IC70：（47.60±4.41）% vs. （31.90 ±4.95）%,t = -6.381,P〈0.05] and72 h [IC50：（52.01±6.63）% vs. （32.65±3.69）%,IC70：（74.31 ±6. 18）% vs. （47.85 ±4.52）%,t = -5. 729
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第10期2388-2391,共4页
Chinese Journal of Experimental Surgery
基金
上海市普陀区肝胆胰重点专科基金项目（B-162）
关键词
蟾毒灵
癌
肝细胞
增殖
凋亡
Bufalin
Cancer, liver cell
Proliferation
Apoptosis
