硒化麒麟菜多糖对膀胱癌细胞生长、凋亡的作用及Bax、Bcl-2、caspase-3蛋白表达的影响

摘要

目的 研究硒化麒麟菜多糖对膀胱癌T24细胞生长、凋亡的作用及Bax、Bcl 2、caspase-3蛋白表达的影响.方法 体外培养T24膀胱癌细胞,分别利用0、0.125、0.25、0.5、1 mg/mL的硒化麒麟菜多糖处理T24细胞24 h、48 h、72 h,利用噻唑蓝(MTT)检测各组细胞增殖情况,0.25、0.5、1 mg/mL的硒化麒麟菜多糖处理细胞48 h后,流式细胞仪检测T24细胞周期分布及细胞凋亡,Western-blot检测T24细胞Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2(Bcl-2)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase-3)蛋白表达.结果 与0 mg/mL处理相比,0.25、0.5、1 mg/mL的硒化麒麟菜多糖处理抑制T24细胞增殖,差异有非常显著性统计学意义(P＜0.01);使用0.125 mg/mL硒化麒麟菜多糖处理,作用24 h时对细胞增殖抑制不显著(P＞0.05),作用48h、72h时对细胞增殖具有较为明显抑制作用(P＜0.05).1 mg/mL的硒化麒麟菜多糖作用后细胞凋亡率高达(23.56±0.96)％.与0 mg/mL处理相比,0.25、0.5、1 mg/mL的硒化麒麟菜多糖作用后,处于G0/G1期的细胞从(52.12±1.74)％分别增加至(64.32±2.42)％、(70.72±1.71)％、(76.09±2.28)％.处于G2/M期和S细胞明显减少(P＜0.05),Bcl-2蛋白的表达明显降低(P＜0.01),Bax、Cleaved-caspase-3蛋白的表达明显升高(P＜0.01).结论 硒化麒麟菜多糖可能通过调控Bax、Bcl-2、Cleaved-caspase-3蛋白表达抑制T24细胞增殖,阻滞细胞周期进程,促进细胞凋亡.%Objective To investigate the effects of seleno-polysaccharide on the growth and apoptosis of bladder cancer cells and on the protein expressions of Bcl-2,Bax and caspase-3.Methods Bladder cancer cell line T24 were cultured in vitro,treated with 0 mg/mL,0.125 mg/mL,0.25 mg/mL,0.5 mg/mL,and 1 mg/mL seleno-polysaccharide for 24 h,48 h and 72 h respectively.The cell proliferation was detected with MTT.After the cells were treated for 48 h,cell cycle and apoptosis were detected with flow cytometry.The protein expressions of Bax,Bcl-2 and caspase-3 were determined with Western blotting.ReSultS Seleno-polysaccharide at concentrations of 0.25 mg/mL,0.5 mg/mL,and 1 mg/mL significantly inhibited cell proliferation (P＜0.01).Seleno-polysaccharide at 0.125 mg/mL for 24 h had no significant inhibitive effect on cell proliferation (P＞ 0.05),while the 48 h and 72 h treatment had a significant inhibitive effect (P＜0.05).The cell apoptosis rate was (23.56± 0.96) ％ after treatment of seleno-polysaccharide at 1 mg/mL.After seleno-polysaccharide treatment at concentrations of 0.25,0.5 and 1 mg/mL,the percentage of cells in G0/G1 phase rose from (52.12 ± 1.74) ％ to (64.32 ± 2.42) ％,(70.72 ± 1.71) ％ and (76.09±2.28)％;percentage of cells in G2/M and S phase decreased significantly (P＜0.05).The protein expression of Bcl-2 significantly reduced,while the protein expressions of Bax and caspase3 significantly increased (P＜0.01).Conclusion Seleno-polysaccharide inhibits T24 cell proliferation,arrests cell cycle,and promotes cell apoptosis by regulating the expressions of Bax,Bcl-2 and caspase-3.