曲马多对体外培养人肝癌细胞株HepG2的影响

The Effect of Tramadol on the Human Liver Carcinoma Cell Line HepG2 in vitro

目的观察曲马多对体外培养人肝癌细胞株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的曲马多(0.1,1,10,100,1000,10000,100000μmol·L-1),对照组RPMI-1640培养液中不加曲马多,分别孵育48小时后,采用MTT比色法检测曲马多对HepG2细胞增殖活性的影响,采用流式细胞技术检测曲马多对HepG2细胞周期的影响,用Hoechst3328染色方法分析鉴定曲马多对HepG2细胞凋亡的影响。结果曲马多浓度≥1000μmol·L-1时,细胞增殖活性较对照组显著下降(P<0.05);随着曲马多浓度的增加,G0/G1期比例逐渐增高,(S+G2)/M期逐渐降低;用Hoechst33258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,曲马多浓度≥10000μmol·L-1时,荧光显微镜下一个视野内凋亡细胞数占细胞总数的比例较对照组显著增加(P<0.05)。结论曲马多在临床常用剂量范围内对人肝癌细胞株HepG2细胞增殖和凋亡无明显影响,浓度≥1000μmol·L-1时可以抑制人肝癌细胞株HepG2增殖,浓度≥10000μmol·L-1时可引起人肝癌细胞株HepG2凋亡。

Objective To evaluate the effects of tramadol on the proliferation, the cell cycle and apoptosis of human liver carcinoma cell line HepG2 in vitro. Methods Human liver carcinoma line HepG2 cells were cultured in vitw. The HepG2 cells of the test group were incubated in the RPMI-1640 medium with tramadol at different concentration（0.1, l, 10, 100, 1000, 10000, 100000μmol.L^-1）. The control gToup were incubated in theRPMI-1640 medium for 48 hours. The level of the cell proliferation was evaluated with MTF method, and the cell cycle was detected with flow cytometry （FCM）, and the morphological changes of apoptosis cells were observed by fluorescence microscopy after staining by Hoechst33258. Results When the concentration of tramadol was over 1000 μmol.L^-1, the level of the proliferation in test group was apparently decreased （P〈0.05）; With the concentra-lion of tramadol increasing, the ratio of G0/G1 phase of HepG2 cells was significantly enhanced and the ratio of S and G/M phase of HepG2 cells was significantly decreased; Under the fluorescent microscopy after staining by the Hoechst33258, it showed that parts of HepG2 cells apoptosis in the test group, the rate of apoptotic cells was significantly increased when the concentration of tramadol was over 10000 μmol.L^-l（p〈0.05）. Conclusion Thedata suggested that tramadol would not inhibit the level of the HepG2 cells proliferation and would not result in apoptosis when the concentration of tramadol was in general range. Tramadol could inhibit the level of the HepG2 cells proliferation when the concentration of tramadol was over 1000μmol.L^-l. Tramadol could result in human liver carcinoma HepG2 cells apoptosis when the concentration of tramadol was over 10000μmol.L^-1.