摘要
目的探讨大黄素对胰腺癌细胞增殖的抑制作用及其机制。方法采用CCK-8法观察不同浓度的大黄素对人类胰腺癌细胞株PC-3细胞增殖的抑制作用;采用荧光显微镜和流式细胞仪观察细胞的凋亡及生长周期的变化;采用透射电镜观察其超微结构的变化;采用免疫组织化学的方法,观察凋亡相关基因蛋白(Fas,Fas-L)表达的变化。结果不同浓度(0、10、20、40和60μmol/L)的大黄素均可抑制PC-3细胞的增殖,而且抑制率呈浓度依赖关系;荧光显微镜检测显示大量早期凋亡细胞(FITC+/PI-),流式细胞仪观察显示明显的凋亡峰出现,并使细胞周期主要被阻滞在G1期(55.19～85.80%);电镜观察显示了大黄素作用后的PC-3细胞具有典型的凋亡细胞形态学特征:细胞膜完整、胞浆浓缩、染色质固缩、核碎裂等;免疫组化研究表明大黄素对PC-3细胞表达的凋亡相关基因具有一定调节作用,即可上调Fas(+～+++)、Fas-L(-～+++)的表达。结论大黄素既能有效地抑制人类胰腺癌PC-3细胞株的生长,又可以诱导癌细胞凋亡,这可能与其能够调节PC-3细胞的Fas和Fas-L凋亡相关基因蛋白的表达有关。
Objective To explore the inhibition effect of emodin on human pancreatic carcinoma cell line PC-3 and its mechanism. Methods CCK-8 assay was used to observe the inhibitory actions of emodin on PC-3 cells at various concentrations. The apoptotic rate and growth cycle of the cells were detected by flow cytometry and fluorescence microscope. The changes of the cells ultrastructures were observed under electron microscope. The expression of apoptosis-related gene protein （Fas,Fas-L） was detected by immunohistochemical staining. Results emodin inhibited the proliferation of PC-3 cells in a concentration-dependent manner. Lots of early apoptotic cell were detected by fluorescence microscope. Marked apoptosis peak was observed and the cells were mainly blocked in G1 phase （55.19～85.80%）. PC-3 cells showed obvious feature of apoptosis under electron microscope,such as intact cell membrane,concentration of plasma,pyknosis of chromatin and nuclear fragmentation. Fas and Fas-L protein were weakly （＋） and negatively （-） expressed in controls respectively,but strongly expressed （＋＋＋） in emodin-treated cells. Conclusion Emodin can not only inhibit the proliferation but also induce the apoptosis of human pancreatic carcinoma cell PC-3.The mechanism is probably related to its effect on the regulation of Fas,Fas-L expression.
出处
《浙江创伤外科》
2010年第3期273-275,共3页
Zhejiang Journal of Traumatic Surgery
基金
浙江省自然科学基金资助项目(Y2080708)
浙江省中医药管理局重点资助项目(2008ZA015)
关键词
大黄素
胰腺癌
CCK-8法
流式细胞术
免疫组化
Emodin
Pancreatic carcinoma
CCK-8 assay
Flow cytometry
Immunohistochemistry
