17-DMAG对肝癌HepG2细胞的作用研究

目的研究HSP90新型抑制剂17-二甲胺乙胺基-17-去甲氧基格尔德霉素(17-dimethy-laminoethylamino-17-demethoxygeldanamycin,17-DMAG)对肝癌细胞株HepG2增殖及凋亡的影响,并对其机制进行初步探讨。方法 MTT比色法检测不同浓度的17-DMAG及5 mg/L的DDP对HepG2细胞的生长抑制率;Annexin V-FITC/PI双染法检测经17-DMAG及DDP作用后HepG2细胞凋亡率的变化。结果 MTT法显示17-DMAG能抑制肝癌HepG2细胞的增殖,且呈时间-剂量依赖性。250 nmol/L的17-DMAG低浓度水平较DDP组对HepG2细胞的抑制率明显增高(P<0.05)。光学显微镜观察17-DMAG作用48 h后HepG2细胞密度减低,细胞体积变小,随着药物浓度增大,细胞数目明显减少。Annexin-FITC/PI双染法检测结果显示,400 nmol/L 17-DMAG干预48 h后细胞早期凋亡率为(22.42±1.83)%,5 mg/L顺铂干预48 h后细胞早期凋亡率为(6.50±1.20)%,未干预组48 h后细胞早期凋亡率为(0.58±0.49)%,表明17-DMAG能诱导HepG2细胞凋亡,且17-DMAG组凋亡率明显高于5 mg/L顺铂组及未干预组(P<0.05)。结论热休克蛋白90抑制剂17-DMAG呈时间-剂量依赖性抑制肝癌HepG2细胞增殖,随着药物浓度的加大,作用时间的延长,细胞增殖能力逐渐下降,并且能诱导HepG2肝癌细胞的凋亡。17-DMAG抑制肝癌细胞的作用较DDP强。

Effect of 17-DMAG on liver cancer cell lines HepG2

【Objective】 To investigate the effect of the novel HSP90 inhibitors 17-DMAG on human liver cancer cell lines HepG2,to see proliferations or apoptosis of HepG2 after drugs and make a preliminary discussion of its mechanisms.【Methods】 MTT assay was applied to measure the optical density of different groups after different concentrations of 17-DMAG and DDP with HepG2 cell line to calculate cell growth inhibition rate and 50% inhibiting concentration(IC50).Optical inverted microscope was used to observe the morphological changes of HepG2 cell line after the drugs.In the end,Annexin V-FITC/PI double staining was to detect apoptosis rate of HepG2 cell line after 17-DMAG and DDP.【Results】 MTT showed 17-DMAG can suppress the proliferation and growth of the HepG2 cell lines.HepG2 cell proliferation inhibition was ascending with an increasing concentration of 17-DMAG and a longer effect time.MTT showed a time-dose effect of a growing inhibition rate of HepG2 cell lines proliferation which is statistically significant(P <0.05).The inhibiting rates of 17-DMAG group above 250 nmol/L level was significantly higher than 5 mg/L DDP group(P <0.05).Along with an increasing concentration of 17-DMAG,what is more,HepG2 cell density was decreased in an optical inverted microscope,the number of HepG2 cells also significantly went down.Flow cytometry analysis showed that apoptosis rate of HepG2 cell line(22.4%) was significantly higher than the normal group(0.58%) after 17-DMAG at an concentration of 400 nmol/L for 48 hours(P <0.01).Compared with the apoptosis rate of DDP group(6.5%),that of the 17-DMAG group has a significant difference,which means 17-DMAG can cause apoptosis of HepG2 cell line more effectively(P <0.01).【Conclusion】 Heat shock protein 90 inhibitors 17-DMAG have significantly inhibition of human liver cancer cell lines HepG2 proliferation which showed a time-dose effect of a growing inhibition rate of HepG2 cell lines proliferation.Along with the increase of drug concentration and the extension of time,the cell proliferation ability gradually declined.17-DMAG can induce HepG2 cell line apoptosis.17-DMAG can exert potent antineoplastic activity more effectively than DDP on human liver cancer cell lines HepG2.