Calphostin C对人视网膜母细胞瘤细胞增殖的抑制作用

Inhibiting effect of Calphostin C on the proliferation of human retinoblastoma oncocytes

目的探讨Calphostin C对人视网膜母细胞瘤(retinoblastoma,Rb)细胞增殖的抑制作用。方法体外培养人Rb44细胞,MTT法检测不同浓度(0.062 5 nmol·L-1、0.125 0 nmol·L-1、0.250 0 nmol·L-1、0.500 0 nmol·L-1、1.000 0 nmol·L-1)Calphostin C作用不同时间(24 h和48 h)对人Rb44细胞增殖的影响;流式细胞术观察不同浓度(0.062 5 nmol·L-1、0.125 0nmol·L-1、0.250 0 nmol·L-1、0.500 0 nmol·L-1、1.000 0 nmol·L-1)Calphostin C作用不同时间(24 h和48 h)对人Rb44细胞周期及凋亡的影响。结果与对照组相比,Calphostin C对体外培养的人Rb44细胞增殖具有显著的抑制作用,并呈一定的浓度和时间依赖性,1.000 0 nmol·L-1Calphostin C作用48 h对人Rb44细胞增殖的抑制作用最明显,增殖抑制率达(70.23±2.43)%。Calphostin C作用人Rb44细胞24 h和48 h的IC50分别为1.742 nmol·L-1和0.258 nmol·L-1。与对照组人Rb44细胞各个细胞周期分布相比,Calphostin C作用24 h后0.500 0 nmol·L-1组和1.000 0 nmol·L-1组G1期细胞比例升高,G2/M期细胞比例明显减少(P<0.05或P<0.01)。Calphostin C作用人Rb44细胞48 h后0.250 0 nmol·L-1组、0.500 0 nmol·L-1组和1.000 0 nmol·L-1组G2/M期细胞比例与对照组相比也明显减少(P<0.05或P<0.01)。与对照组人Rb44细胞凋亡率相比,除0.062 5 nmol·L-1Calphostin C作用24 h组外,其余各组人Rb44细胞的凋亡率均显著升高,差异均有统计学意义(P<0.05或P<0.01),且随着Calphostin C浓度的增加,细胞凋亡率增高,呈一定的剂量依赖性。结论 Calphostin C对Rb细胞的体外增殖具有一定的抑制作用,同时对细胞凋亡起着促进作用,能够阻滞细胞周期进程。

Objective To explore the inhibiting effect of Calphostin C on the proliferation of human retinoblastoma （Rb） cells. Methods Human Rb44 cells were cultivated in vitro. MTT and flow cytometry （FCM） were performed respectively to measure the influence of Calphostin C（0.052 5 nmol ·L^-1 ,0. 125 0 nmol ·L^-1 ,0.250 0 nmol ·L^-1 ,0.500 0 nmol·L^-1,1. 000 0 nmol ·L^-1 ） on the proliferation of human Rb44 cells after different action time（24 hours,48 hours）. Results Compared with the con- trol, Calphostin C was of great inhibiting effect on the proliferation human Rb44 cells in a certain concentration-and time-dependent manner. Calphostin C（ 1. 000 0 nmol · L^-1 ） with an action time of 48 hours had the strongest inhibiting effect, the proliferation inhi- bition rate was （70.23 ＋ 2.43 ） %. Calphostin C IC50 on human Rb44 cells were 1. 742 nmol ·L^-1 and 0.258 nmol ·L^-1 when action time were 24 hours and 48 hours reapec- tively. Compared with distribution of cell cycles of the control, the proportion of G1 phase cells increased, while that of the G2/M phase cells decreased significantly in Cal- phostin C groups （0. 500 0 nmol ·L^-1, 1. 000 0 nmol ·L^-1 ） with an action time of 24 hours（P 〈 0. 05 or P 〈 0.01 ）. G2/M cells in Calphostin C groups（0. 250 0 nmol ·L^-1, 0. 500 0 umol ·L^-1,1. 000 0 umol ·L^-1 ） with an action time of 48 hours were less thanthat of the control（P 〈 0.05 or P 〈 0.01 ）. Apoptosis rate of human Rb44 cells of all groups were significantly higher than that of the control, except for the Calphostin C group （0.052 5 nmol ·L^-1 ） with an action time of 24 hours （P 〈 0.05 or P 〈 0.01 ）. The apoptosis rate increased with the rise of Calphostin C concentration, which showed a certain dose-dependent manner. Conclusion Calphostin C has a certain inhibiting effect on the proliferation of human Rb cells in vitro, and it could promote human Rb cell apoptosis and impede the process of cell cycles.