[Study of effects of micheliolide on ovarian cancer cell lines and its mechanism].
Objective:To study the roles of micheliolide on ovarian cancer cells.Methods:Firstly, human ovarian cancer cell lines HeyA8, SKOV3 and A2780/DDP were treated with different concentration of micheliolide(0.25, 0.5, 1, 2.5, 5, 10, 20, 50 μmol/L)for 72 hours, then methyl thiazolyl tetrazolium(MTT)assay was used wo detect the growth of the human ovarian cancer cell lines and the stongest inhibited cell line were selected for the following test. Secondly, after HeyA8 cell line was treated with different concentration(5, 10, 20 μmol/L)of micheliolide for 24 hours, the HeyA8 cell apoptosis was measured byflow cytometry. Thirdly, the expression of RelA mRNA in HeyA8 cell was detected through real-time PCR, the expressions of nuclear factor κB(NF-κB)signal pathway related protein RelA and the activited cysteinyl aspartate specific proteinase(caspase-9)were detected by western blot analysis.Results:(1)The growth of HeyA8, SKOV3 and A2780/DDP cells were all significantly inhibited after being treated with different concentration of micheliolide for 72 hours and the roles of inhibition were all concentration dependant(P<0.05). The half maximal inhibitory concentration(IC50)of HeyA8, SKOV3 and A2780/DDP were(9.8±2.2),(12.0±2.1)and(12.8±1.8)μmol/L, respectively. We chose HeyA8 cell to do the following expreriments because of its best inhibited effect.(2)After HeyA8 cell was treated with micheliolide of different concentrations, as the concentration increased(20 and 0 μmol/L, for example), the apoptosis rate of HeyA8 cell raised from(7.2±1.0)% to(17.4±1.1)%, the percentage of survived cells reduce from(92.8 ± 1.3)% to(82.6 ± 1.4)%, and the relative mRNA level of RelA decreased from 1.00 ± 0.13 to 0.18 ± 0.00(P<0.01); furthermore, the expression of RelA protein was weaken and the activited caspase-9 protein expression was increased gradually.Conclusions:Micheliolide plays a significantly inhibited role in HeyA8, SKOV3 and A2780/DDP cells. The inhibited role of micheliolide inovarian cancer cells might through inhibiting nuclear factor-kappa B(NF-кB)signaling pathway, and inducing the expression of activited caspase-9 protein to promoting apoptosis of HeyA8 cell.
Department of Obstetric and Gynecology, Tianjin Medical University General Hospital, Tianjin 300052, China.
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Li
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Ding
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Ji
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Gao
D017209:Apoptosis; D053453:Caspase 9; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019008:Drug Resistance, Neoplasm; D005260:Female; D006801:Humans; D016328:NF-kappa B; D010051:Ovarian Neoplasms; D060888:Real-Time Polymerase Chain Reaction; D045789:Sesquiterpenes, Guaiane; D051996:Transcription Factor RelA
D016328:NF-kappa B; C498143:RELA protein, human; D045789:Sesquiterpenes, Guaiane; D051996:Transcription Factor RelA; C577928:micheliolide; C506433:CASP9 protein, human; D053453:Caspase 9