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P-Methoxycinnamaldehydelevofloxacin-3-Ylhydra Zone Ininduces Apoptosis of Human Hepatocarcinoma SMMC-7721 Cells

Author: WangDeHui
Tutor: LiuBin
School: Henan University
Course: Biochemistry and Molecular Biology
Keywords: Quinolone derivatives Hepatoma cells Cell cycle Apoptosis Mitochondrial membrane potential
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 15
Quote: 0
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Abstract


Objective: 10 quinolone derivatives as candidate agents were screened for find the compounds which have high inhibiting activity to proliferation of SMMC-7721 cells, and studied the antineoplastic mechanism of the compounds in order to provide references for the development of new anticancer drugs.Methods: The proliferation of the cells and the inhibition effect of quinolone derivatives on the cell proliferation were examined by MTT assay. PI fluorescence staining with flow cytometric analysis was used to analysis cell cycle. Cell apoptosis was observed by fluorescence microscope techniques with Hoechst 33258 associated with propidium iodide （PI） staining and TUNEL assay. DNA ladder was observed through agarose gel electrophoresis. Mitochondrial membrane potential（△ψm） were measured by high content screening. The caspase-9, caspase-8, caspase-3, p53, Bcl-2 and Bax protein expressions were detected by western blot analysis.Results: 10 fluoroquinolones hydrazone compounds （FQ1 FQ10） screening results showed that cell proliferation. The drug screening indicated that FQ3、FQ8、and FQ10 possessed high inhibiting activity to proliferation of SMMC-7721 cells. Compared to the control, the inhibitory rate of cell proliferation is significant difference with FQ10 of 0.625μmol·L-1 at 24 h（p<0.05）; While the significant difference gets more significant with FQ10 of （ 0.62510）μmol·L-1 at 24 h（p<0.01）.FQ10 inhibited the proliferation of SMMC-7721 cells with the IC50 of 4.40μmol·L-1at 24 h in a dose- and time- dependent manner; Cell cycle analysis showed that, with increasing concentration FQ10, G0/G1 phase decreased, S phase increased, cell cycle arrest at G2 / M phase; Fluorescence staining showed morphological changes, different concentrations of FQ10 role in cells, apoptotic cells increased and visible chromatin condensation, nuclear fragmentation into pieces and other typical characteristic changes of apoptosis. TUNEL assay indicate that the apoptosis rate increased along with the concentration of FQ 10 in a dose-dependent manner, Experimental statistics of the drug treatment group higher apoptosis rate （P <0.05）; Characteristic morphological changes of apoptotic cells can be seen clearly. Typical DNA ladder bands have indicated that multiple apoptotic cells upon 10μmol·L-1 FQ10 incubation with SMMC-7721 cells for 24h. Western blot results showed, Expression of apoptosis related proteins p53, Bax, caspase-9 and caspase-3 increased significantly compared to the control, at the same time expression of Bcl-2 decreased. caspase-8 protein expression did not change significantly.Conclusions: FQ10 play a potential role in inhibiting the proliferation of SMMC-7721 cells and arrest cell cycle in a dose- and time- dependent manner. meanwhile, FQ10 induced apoptosis of SMMC-7721 cells in a mechanism related to mitochondrial apoptosis pathway.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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