摘要
目的了解阿片类镇痛药瑞芬太尼对体外培养肝癌细胞株HepG2细胞增殖、细胞周期及细胞凋亡的影响。方法体外培养人肝癌HepG2细胞，实验组在RPMI-1640培养液中加入不同浓度瑞芬太尼（0．001、0．01、0．1、1、10、100、200μmol／L），对照组RPMI-1640培养液中不加瑞芬太尼，分别孵育48h后，采用MTT比色法检测HepG2细胞增殖活性，采用流式细胞技术检测瑞芬太尼对HepG2细胞周期的影响，用Hoechst33258染色方法分析鉴定瑞芬太尼对HepG2细胞凋亡的影响。结果瑞芬太尼对HepG2细胞增殖的抑制作用呈剂量依赖性，瑞芬太尼浓度≥1μmol／L开始，HepG2细胞增殖较对照组显著下降（P〈0．05），但在100μmol／L与200μmol／L浓度时细胞增殖改变差异无统计学意义（P〉0．05）；瑞芬太尼浓度≥1μmol／L时，G0／G1期细胞逐渐明显多于对照组（P〈0．05），S期细胞减少（P〈0．05），使HepG2细胞阻滞于G0／G1期；用Hoechst33258方法染色后在表面荧光显微镜下发现实验组部分HepG2细胞发生凋亡；荧光显微镜下一个视野内凋亡细胞占总细胞数的比例较对照组显著增加（P〈0．05）。结论当瑞芬太尼浓度≥1μmol／L时，能够抑制肝癌细胞株HepG2细胞增殖并诱导其凋亡。
Objective To evaluate the effects of remifentanil on the proliferation, the cell cycle and apoptosis of human liver carcinoma cell line HepG2 in vitro. Methods Human liver carcinoma cells HepG2 were cultured in vitro. The HepG2 cells of the test group were incubated in the RPMI-1640 medium with remifentanil at different concentration（0. 001,0. 01,0. 1,1,10,100,200 μmol/L）. The HepG2 cells of the control group were incubated in the RPMI-1640 medium for 48 hours. The level of the cell proliferation was evaluated with methyhhiazolyldiphenyl - tetrazolium bromide （MTF） method. The cell cycle was detec- ted with flow cytometry （FCM）. The morphological change of apoptosis cell was observed by fluorescence microscopy after staining by Hoechst33258. Results Remifentanil inhibited the proliferation of the HepG2 cells with a dose-dependent effect. Compared with control group, the cell proliferation capability was appar- ently decreased in the test group （ P 〈 0. 05 ） when the concentration of remifentanil was over 1 μmol/L （ P 〈0. 05）. However, no significant difference in cell proliferation was found when remifentanil was 100 and 200 μmol/L. The ratio of G0/G1 phase of HepG2 cells was significantly enhanced and the ratio of S phase of HepG2 cells was significantly decreased when remifentanil was over 1μmol/L. The fluorescent mi- croscopy stained by the Hoechst33258 showed part of HepG2 cells apoptosis in test group, and the apoptotic rate was significantly increased when remifentanil was over 1μmol/L （ P 〈 0.05 ）. Conclusions The datasuggest that remifentanil would inhibit the proliferation of HepG2 cells and induce apoptosis when remifen- tanil was over 1 μmol/L.
出处
《中国医师杂志》
CAS
2013年第5期577-580,共4页
Journal of Chinese Physician
基金
国家自然科学基金（21135001）,中国博士后科学基金（201104503）,湖南省科技计划项目（2011RS4038）,湖南省肿瘤医院科研平台建设（PT2012-5）
关键词
芬太尼
药理学
肝肿瘤
病理学
肿瘤细胞
培养的
细胞增殖
细胞凋亡
Fentanyl/pharmacology
Liver neoplasms/pathology
Tumor cells, cultured
Cell pro- liferation
Apoptosis
