Trimethoxy-benzaldehyde ciprofloxacin schiff base induces apoptosis in human hepatocarcinoma SMMC-7721 cells / 中国药学杂志

ABSTRACT

OBJECTIVE: To investigate the effect of 1-cyclopropyl-6-fluoro-7-(piperazin-1-yl)-3-[5-benzylsulfanyl-4-(3, 4, 5-trimethoxybenzylidene)amino-4H-1, 2, 4-triazol-3-yl]-quinolin-4(1H) -one (M27) on apoposis in hepatocarcinoma SMMC-7721 cells in vitro. METHODS: SMMC-7721 cells, colon adenocarcinoma cells(HCT-116) and leukemia cell line JURKET were treated by M27 with different concentrations for different time in vitro, the inhibitory effect of M27 and its precursor ciprofloxacin on the cell proliferation were examined by MTT assay. Cell apoptosis was determined by Hoechst 33258 fluorescence staining and TUNEL assay. The effect of M27 on topoisomerase II activity was measured using agarose gel electrophoresis by Plasmid pBR322 DNA as the substrate. Mitochondrial membrane potential(Δψm) was measured by high content screening imaging system. The p53, Caspase-9, Caspase-3, Caspase- 8, Bcl-2, Bax and cytochrome C protein expressions were determined by Western blotting analysis. RESULTS: The proliferation of the cancer cells was inhibited by M27 at 10-60 μmol·L-1 in time-and dose-dependent manner. Ciprofloxacin showed weak cytotoxicity against SMMC-7721 cell. SMMC-7721 cells treated by M27 with different concentrations for 24 h increased the percentage of apoptosis cells obviously (P < 0.05) with a decrease in the mitochondrial membrane potential. Compared with control group, M27 influenced obviously DNA topoisomerase II activity, stimulated DNA cleavage and inhibited DNA reunion mediated by topoisomerase II. In addition, M27 increased protein expression of p53, Bax, Caspase-8, Caspase-9, Caspase-3, as well as the cleaved activated forms of Caspase-9, Caspase-8 and Caspase-3 significantly, whereas the expression of Bcl-2 decreased. There was a significant increase of cytochrome C in the cytosol after 24 h of treatment with M27 and a decrease in the mitochondrial compartment. CONCLUSION: M27 as a fluoroquinolone derivative exerted potent anticancer activity through the mechanism of eukaryotic topoisomerase II poisoning. The growth inhibition is mainly mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.