三氧化二砷注射液对胰腺癌细胞系PC-3的体外作用

Effect of arsenic trioxide on pancreatic carcinoma cell line PC-3 in vitro

目的:探讨三氧化二砷(亚砷酸,As2O3)注射液对人类胰腺癌细胞的生物学效应及其作用机制. 方法:采用MTT法观察不同浓度的As2O3对人类胰腺癌细胞株PC-3细胞生长的抑制作用;以流式细胞术观察细胞的凋亡率及生长周期的变化;以透射电镜观察其超微结构的变化;以免疫组织化学的方法,观察凋亡相关基因蛋白(Fas,Fas—L,Bcl-2和Bax)表达的变化. 结果:不同剂量(1、2和4 mg/L)的As2O3均可抑制PC- 3细胞的增殖,且抑制率具有浓度和时间依赖性;流式细胞仪检测显示明显的凋亡峰出现,并使细胞周期主要被阻滞在S期(14.86—63.66%);电镜观察显示了药物作用后的癌细胞具有明显的凋亡特征性改变:细胞膜完整、染色质固缩、核碎裂、凋亡小体形成等;免疫组化研究表明As2O3注射液对PC-3细胞表达的凋亡相关基因具有一定调节作用,即可下调Bcl-2 (+++～+)的表达,上调Fas(+～+++)、Fas-L(-～+++) 的表达. 结论:As2O3注射液对人类胰腺癌细胞株PC-3具有较强的直接抑制作用,可以诱导癌细胞凋亡,这可能与其能够调节癌细胞的Fas,Fas—L和Bcl-2等基因蛋白的表达密切相关.

AIM： To explore the biological effect of arsenic trioxide （As2O3） on human pancreatic carcinoma cell line PC-3 and its mechanism. METHODS： MTT assay was used to observe the inhibitory actions of As2O3 on PC-3 cells at various concentrations. The apoptotic rate and growth cycle of the cells were detected by flow cytometry. The changes of the cells＇ ultrastructures were observed under electron microscope. The expression of apoptosis-related gene protein （Fas,Fas-L, Bcl-2, Bax） was detected by immunohistochemical staining.RESULTS： As2O3 inhibited the proliferation of PC-3 cells in a concentration- and time-dependent manner. Marked apoptosis peak was observed and the cells were mainly blocked in S phase （14.86-63.66%）. PC-3 cells showed obvious feature of apoptosis under electron microscope, such as intact cell membrane, pyknosis of chromatin, nuclear fragmentation and apoptotic body formation. Bcl-2 protein was weakly expressed （＋＋＋） in controls, but strongly expressed （＋） in As2O3-treated cells. Fas-L protein were weakly （＋） and negatively （-） expressed in controis respectively, but strongly expressed （＋＋＋） in As2O3-treated cells. CONCLUSION： As2O3 can inhibit the proliferation and induce the apoptosis of human pancreatic carcinoma cell PC-3 and the mechanism is probably related to its effect on the regulation of Fas, Fas-L and Bcl-2 expression.