环孢素A与他克莫司对人肝癌HepG-2细胞生物学行为的影响

Influence of cyclosporine A and tacrolimus on hepatocellular carcinoma HepG-2 cell line biological behaviors

目的探讨环孢素A(CsA)与他克莫司(FK506)对肝癌的增殖、凋亡、复发及转移的影响。方法培养人肝癌细胞HepG-2,分别经5、10、100、500、1 000、5 000μg／L的CsA、FK506处理24～48 h后,采用噻唑蓝(MTT)法检测细胞增殖、流式细胞术测定细胞周期及细胞凋亡、荧光定量聚合酶链反应(PCR)检测转化生长因子(TGF)-β、细胞间黏附分子(ICAM)、骨桥蛋白(OPN)。结果肝癌细胞增殖能力48 h明显受到抑制．CsA、FK506使G_2期显著延长;FK506在小于100 μg／L及CSA在小于1 000μg／L时,随浓度增加,肝癌凋亡率逐渐增加,当两者分别超过100、1 000 μg／L时,则出现凋亡率减低;FK506、CSA分别在100、1 000μg／L低浓度时对OPN无影响,在 1 000、5 000μg／L高浓度时基因量显著增加。FK506在低浓度时对ICAM及TGF-β无影响,在高浓度时基因量显著增加;而CSA对两者无明显影响。结论 CsA、FKS06均对肝癌增殖有抑制作用,使G_2期阻滞,促凋亡;低浓度时对肝癌转移无影响,高浓度时增加转移危险。

Objective To investigate the effect of cyclosporine A （CsA） and tacrolimus （FK506） on the growth and metastasis of hepatocellular carcinoma （HCC）. Methods Human HCCcell line HepG- 2 was cultured at different concentrations of CsA and FK506 respectively for 24 to 48 h. Cell growth was assessed with MTT assay, cell cycle and cell apoptosis were assessed by flow cytometry, and transforming growth factor hetal （ TGF-β）, intercellular adhesion molecule （ ICAM）, osteopontin （OPN） were analyzed by fluorescent quantitative polymerase chain reaction （PCR）. Results The ability of growth was inhibited in 48 h. CsA and FKS06 could increase cell ratio of G2 phase. The apoptosis ratio was increased when FK506 was below 100 μg/L and CSA was below 1 000μg/L, and when they were beyond the concentration, the apoptosis ratio was decreased. FK506 and CSA had no influence on OPN mRNA when they were at 100, 1 000 μg/L concentration, and increased OPN mRNA when they were at 1 000, 5 000 μg/L concentration respectively. FK506 influenced the ICAM and TGF-β in the same way, and CsA has no influence on them. Conclusion CsA and FK506 could inhibit the growth of HCC cells, arrest G2 phase and increase apoptosis. CsA and FK506 have no influence on metastasis of HCC when FKS06 was below 100 μg/L and CSA was below 1 000 μg/L, and could increase the risk of metastasis when they were beyond the concentration.