摘要
目的 研究异喹啉生物碱克班宁对肝癌细胞株Huh7的生长抑制作用及促凋亡作用。方法用不同浓度的克班宁作用于Huh7细胞后,按照CCK-8法检测克班宁对肝癌细胞增殖的影响,并据此将后面的实验分为对照组和给药组(36、48、60μg·mL^(-1));利用形态学实验观察克班宁对肝癌细胞生长和状态的影响,平板克隆实验观察细胞克隆形成能力;Hoechst 33258实验和流式细胞术检测细胞凋亡状态和水平;Western blot检测细胞中BAX、Bcl-2、PARP凋亡相关蛋白及AKT、p-AKT、FoxO3a、p-FoxO3a的表达情况。结果 与对照组相比,克班宁可明显抑制Huh7细胞的增殖能力,抑制作用呈剂量和时间依赖性;且可诱导Huh7细胞出现明显的形态学变化;并在长时间内影响其克隆形成能力;同时还显著促进细胞凋亡,上调PARP、BAX蛋白的表达,下调Bcl-2、p-AKT、p-FoxO3a蛋白的表达。结论 克班宁不仅能抑制肝癌细胞株Huh7的增殖,还能促进细胞凋亡,且其促凋亡作用有可能是通过抑制AKT/FoxO3a信号通路产生的。
Objective To determine the inhibitory effect of isoquinoline alkaloid crebanine on the proliferation and apoptosis of hepatocellular carcinoma cell line Huh7. Methods After different concentrations of crebanine acting on Huh7 cells, the effect of crebanine on the proliferation of Huh7 cells was detected according to CCK-8 method. The following experiments were divided into a control group and 3 dosing groups(36, 48 and 60 μg·mL^(-1));subsequent morphological experiment was canducted to determine the effect of crebanine on the growth and status of Huh7 cells. The ability of cell clone formation was observed in plate cloning assays. Then, Hoechst 33258 assay and flow cytometry were used to detect the apoptosis status and level. The expression of apoptosis-related proteins BAX, Bcl-2, PARP, AKT, p-AKT, Fox O3a, and p-Fox O3a in the cells were recognized by Western blot. Results Compared with the control group, crebanine significantly inhibited the proliferation of Huh7 cells with dose and time dependence. It evidently altered the morphology of Huh7 cells and affected their reproduction over an extended period of time. Crebanine also promoted the apoptosis significantly, up-regulated PARP and BAX protein expression, and down-regulated Bcl-2, p-AKT, and p-FoxO3a protein expression. Conclusion Crebanine not only inhibits the proliferation of hepatocellular carcinoma cell line Huh7, but also promotes the apoptosis by inhibiting AKT/FoxO3a signal pathway.
作者
谭佳杰
向玉玲
熊远果
张洪
TAN Jia-jie;XIANG Yu-ling;XIONG Yuan-guo;ZHANG Hong(Department of Pharmacy,Renmin Hospital of Wuhan University,Wuhan 430060)
出处
《中南药学》
CAS
2023年第2期392-397,共6页
Central South Pharmacy
基金
湖北省自然基金项目(No.2012FKB04439)。
