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机构地区:[1]江西省妇幼保健院病理科,南昌330006 [2]中南大学湘雅医学院病理学教研室,长沙410000
出 处:《江西医药》2016年第10期1023-1027,共5页Jiangxi Medical Journal
摘 要:目的观察碱性成纤维细胞成长因子(FGF-2)对小细胞肺癌(SCLC)细胞株NCI-H446中凋亡抑制蛋白Survivin蛋白的表达水平和细胞凋亡的影响,以及探讨PKC对此过程的调控作用。方法采用Western blot检测FGF-2对NCI-H446细胞中Survivin蛋白、细胞总蛋白和膜蛋白中PKCα的表达水平的变化;特异性PKC抑制剂Calphostin C抑制PKC活性,Western blot检测NCI-H446细胞中Survivin蛋白、细胞总蛋白和膜蛋白中PKCα的表达水平的变化;用流式细胞术和Hoechst 33258荧光染色两种方法检测FGF-2作用后及Calphostin C干预后细胞凋亡率的改变。结果 FGF-2能上调NCIH446细胞Survivin蛋白的表达(50μg/l FGF-2作用NCI-H446细胞4h后实验组Survivin蛋白表达显著高于对照组);FGF-2快速激活NCI-H446细胞中PKC信号通路(50μg/l FGF-2作用NCI-H446细胞后,PKCα总蛋白无变化,PKCα膜蛋白表达增加,并在刺激30min时达到峰值,随后开始下降);FGF-2能通过PKC信号通路上调NCI-H446细胞Survivin蛋白的表达;FGF-2能通过PKC信号通路抑制NCI-H446细胞的凋亡[利用流式细胞术和Hoechst 33258染色两种方法检测空白组、刺激组、阻断组各组NCI-H446细胞凋亡率,结果分别为(12.3±0.004)%,(6.7±0.006)%,(10.37±0.014)%及(6.15±0.8)%,(3.89±0.6)%,(5.11±1.0)%,各组之间差异有统计学意义(P<0.05)]。结论 FGF-2可通过PKC信号传导途径来促进小细胞肺癌NCIH446细胞Survivin蛋白的表达,从而抑制细胞凋亡。Objective To investigate the effect of FGF-2 on the explanation of Survivin and the apoptosis in SCLC NCI-H446 cells,and to elucidate the role of PKC in this effect.Methods Western blot was used to detect the expression of Survivin protein and the PKα protein induced by FGF-2 in NCI-H446 cells;The NCI-H446 cells were pretreated with Calphostin C,then were treated by FGF-2,then the survivin proterin and the PKCα proterin were detected by Western blot,and then the respective apoptosis were detected by flow cytometry and Hoechst 33258 stainng.Resluts FGF-2 increased the expression of Survivin in the NCI-H446 cells(when treated with 50μg/l FGF-2 for 4h,the expression of Survivin became obviously increased compared with blank group);FGF-2 actived the PKC signaling pathway in NCI-H446 cells quickly(the expression of PKCα proterin in the membrane of the NCI-H446 cells reached the maximum level when cultured with 50μg/l FGF-2 for 30 min,but after that they became decreased);FGF-2 induced the expression of Survivin through the PKC signaling pathway;FGF-2 could inhibit the apoptosis of the NCI-H446 cells through the PKC signaling pathway [two methods including flow cytometry and Hoechst 33258 stainng in the blank,stimulation and block groups showed the NCI-H446 cells apoptosis rate were(12.3 ±0.004)%,(6.7 ±0.006)%,(10.37 ±0.014)% and(6.15 ±0.8)%,(3.89 ±0.6)%,(5.11 ±1.0)%,with significant differences between the groups(P〈0.05)].Conclusions FGF-2 could inhibit the cells apoptosis by inducing the explanation of survivin in NCI-H446 cells,PKC may play an important role in this process.
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