[The cytotoxicity of indirubin derivative PHII-7 against human breast cancer MCF-7 cells and its mechanisms].

2012 
Objective To observe the cytotoxicity of indirubin derivative PHⅡ-7 against human breast cancer MCF-7 cells and to study its primary mechanisms. Methods The proliferation of MCF-7 cells was detected using MTT colorimetry. Annexin Ⅴ/PI double staining was applied to detect the apoptosis rate of MCF-7 cells. The distribution of cell cycles was detected using PI staining and flow cytometry (FCM). The levels of reactive oxygen species (ROS) in MCF-7 cells were detected by DCFH-DA staining. The mRNA and protein levels of c-fos were detected using RT-PCR and Western blot analysis. Results PHⅡ-7 at different concentrations inhibited the proliferation of MCF-7 cells in a concentration-dependent manner, with the inhibitory rate ranging from 43.13% to 90.90% (P0.05). The inhibition was strengthened along with increased concentrations. PHⅡ-7 at different concentrations could induce the apoptosis of MCF-7 cells. The early apoptosis rate was 1.43% ± 0.02%, 9.14% ± 0.36%, and 45.79% ± 8.46%, respectively with the action of 1.25, 2.50, and 5.00 μmol/L PHⅡ-7, respectively, showing dose-dependent manner. FCM analysis found that the proportion of MCF-7 cells in the G0/G1 phase and the S phase decreased after treatment with PHⅡ-7, and the ratio of MCF-7 cells in the G2/M phase obviously increased (P0.01). The intra-cellular ROS level was significantly elevated 2 h after pretreatment with PHⅡ-7. The levels of the protooncogene c-fos mRNA and protein were down-regulated in a dose-dependent manner after action of PHⅡ-7. Conclusions PHⅡ-7 exerted obvious in vitro cytotoxic effects on MCF-7 cells. Its mechanisms might be associated with arresting the cell cycle, regulating the redox equilibrium, and down-regulating the expression of the protooncogene.
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