摘要
目的研究海兔素(aplysin)对人乳腺癌FCM-7细胞的抑制作用,并探讨其可能的作用机制。方法 MTT法测定海兔素对人乳腺癌FCM-7细胞增殖的影响;流式细胞术测定细胞凋亡情况、细胞周期变化和增殖细胞核抗原(PCNA)、Fas、bc1-2阳性细胞百分率及细胞基质钙离子(IE Ca2+)含量变化。结果经10、20、40、60、80 mg/L海兔素处理FCM-7细胞48h后,细胞的生长增殖明显受到抑制,呈量效依赖性,其IC25和IC50值为分别为28.3和31.9 mg/L;经10和40 mg/L海兔素处理FCM-7细胞48h,出现明显的凋亡亚二倍体峰,细胞凋亡率分别为5.08%,和33.6%,且随着海兔素剂量不断增加,G0/G1期细胞所占比例也逐渐升高,而S期细胞所占比例则呈逐渐下降趋势,其中,40 mg/L海兔素组与对照组及10 mg/L海兔素组比较,差异有统计学意义(P<0.05)。经10、20、40 mg/L海兔素作用于FCM-7细胞,其Fas蛋白表达阳性细胞百分率均明显升高,分别达到(14.61±1.23、25.65±1.57、38.65±1.89)%,与对照组比较及各药物组间比较,差异有统计学意义(P<0.05);而PCNA及Bc1-2蛋白表达则逐渐下降,其中,20、40 mg/L海兔素组Bc1-2和PCNA表达均较对照组降低,且海兔素浓度越高,Bc1-2和PCNA表达越低,经统计学处理,差异有统计学意义(P<0.05)。经10、20、40 mg/L海兔素作用于FCM-7细胞,IECa2+含量也显著升高,与对照组及海兔素各组间比较,差异有统计学意义(P<0.05)。结论海兔素可诱导乳腺癌FCM-7凋亡,其作用机制可能与aplysin阻滞FCM-7细胞周期,同时提高Fas促凋亡蛋白表达和Ca2+释放,抑制PCNA和Bc1-2等细胞增殖相关蛋白表达有关。
Objective To investigate the effects of aplysin on the apoptosis of human breast cancer FCM- cells in vitro. Methods The apoptosis of FCM- cell lines was determinated by MTT assay. Cell cycle and positive rate of proliferation cell nuclear antigen （PCNA）, apoptosis associated protein Fas and be 1- and intracelular calcium ions （IE Ca^2- levels were measured by flow cytometry. Results Aplysin could decrease the proliferation of FCM- cells significantly in a dose-ependent manner. The IC25 and ICs0 of aplysin on FCM-cell for 48h were 28.3 and 31.9 mg/L. When treating FCM- with aplysin at concentrations of 10, 20, 40, 60, 80 mg/L for 48 h, the growth of the cells was obviously inhibited. Typical apoptosis sub-iploid peak was found when the concentration of aplysin was 10 and 40 mg/L, and the apoptotie rates of FCM- was 5.08% and 33.6% respectively, which was significantly higher than 1.83% in control group （P〈0.05）. Cellular quantity increased from （59.3±.27）% to （79.54±37）% at G0/G1 phase, respectively, which was significantly higher than （48.64±92）% in control group. Meanwhile, cellular quantity decreased from （27.3±63）% to （15.0±34）% at S phase, respectively, which was significantly lower than （31.5±72）% in control group （P〈0.05）. Cell cycle was arrested in G0/GI phase. Compared with control group, aplysin could effectively increase Fas expression and IECa- levels, and inhibited theexpressions of PCNA and Bcl- significantly in FCM- cells （P〈0.05）. Conclusion Aplysin can suppress the proliferation and induces apoptosis of FCM- cells by increasing the expression of Fas and inhibiting the expression of PCNA and bcl- [ACTA NUTRIMENTA SINICA, 2013,35（6）： 599-6031
出处
《营养学报》
CAS
CSCD
北大核心
2013年第6期599-603,共5页
Acta Nutrimenta Sinica
基金
山东省科技厅科技攻关项目(No.2006GG2302002)
