Limits...
Trimethoxy-benzaldehyde levofloxacin hydrazone inducing the growth arrest and apoptosis of human hepatocarcinoma cells.

Sun JP, Shi ZY, Liu SM, Kang YH, Hu GQ, Huangfu CS, Deng JB, Liu B - Cancer Cell Int. (2013)

Bottom Line: There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment.In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, 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.The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Nursing, Institute of Neurobiology, Henan University, Kaifeng, China. kangyh0811@gmail.com.

ABSTRACT

Background: In order to search for new structural modification strategies on fluoroquinolones, we have designed and synthesized a series of fluoroquinolone derivatives by linking various hydrazine compounds to the C-3 carboxyl group of levofloxacin and assessed their anticancer activities. Several novel levofloxacin derivatives displayed potent cytotoxicity against the tested cancer cell lines in vitro. In the present study, we investigated the effect of 1-Cyclopropyl-6-fluoro-4-oxo-7- piperazin-1, 4-dihydro- quinoline- 3-carboxylic acid benzo [1,3] dioxol-5- ylmethylene- hydrazide (QNT11) on the apoptosis of human hepatocarcinoma cells in vitro.

Methods: The inhibition effects of QNT11 on cell proliferation were examined by MTT assay. Cell apoptosis was determined by TUNEL and DNA agarose gel electrophoresis method. The topoisomerase ΙΙ activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Cell cycle progression was analyzed using flow cytometry in conjunction with ethanol fixation and propidium iodide staining. Mitochondrial membrane potential (△ψm) was measured by high content screening image system. The caspase-9, caspase-8, caspase-3, Bcl-2, Bax, CDK1, Cyclin B1and cytochrome c protein expressions were detected by Western blot analysis.

Results: QNT11 showed selective cytotoxicity against Hep3B, SMMC-7721, MCF-7 and HCT-8 cell lines with IC50 values of 2.21 μM, 2.38 μM, 3.17 μM and 2.79 μM, respectively. In contrast, QNT11 had weak cytotoxicity against mouse bone marrow mesenchymal stem cells (BMSCs) with IC50 value of 7.46 μM. Treatment of Hep3B cells with different concentrations of QNT11 increased the percentage of the apoptosis cells significantly, and agarose gel electrophoresis revealed the ladder DNA bands typical of apoptotic cells, with a decrease in the mitochondrial membrane potential. Compared to the control group, QNT11 could influence the DNA topoisomerase IIactivity and inhibit the religation of DNA strands, thus keeping the DNA in fragments. There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment. Observed changes in cell cycle distribution by QNT11 treated might be caused by insufficient preparation for G2/M transition. In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, 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.

Conclusions: Our results showed that QNT11 as a fluoroquinolone derivative exerted potent and selectively anticancer activity through the mechanism of eukaryotic topoisomerase II poisoning. The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

No MeSH data available.


Related in: MedlinePlus

Effects of QNT11 on the mitochondrial membrane potential of Hep3B cells HCS analysis after JC-1 staining to detect changes in the mitochondrial membrane potential of Hep3B cells Induced by QNT11. Cells were treated with 0, 0.803 (IC30 group), 2.210 (IC50 group)and 3.616 μM (IC70 group) QNT11 respectively for 24 h. The mitochondrial membrane potential (△ψm) was measured by HCS image system.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3716872&req=5

Figure 5: Effects of QNT11 on the mitochondrial membrane potential of Hep3B cells HCS analysis after JC-1 staining to detect changes in the mitochondrial membrane potential of Hep3B cells Induced by QNT11. Cells were treated with 0, 0.803 (IC30 group), 2.210 (IC50 group)and 3.616 μM (IC70 group) QNT11 respectively for 24 h. The mitochondrial membrane potential (△ψm) was measured by HCS image system.

Mentions: Mitochondrial membrane potential (△ψm) was detected with the fluorescent probe JC-1, which exists predominantly in monomeric form in cells with depolarized mitochondria and displays green fluorescence at 490 nm. On the other hand, JC-1 primarily forms aggregates in cells with polarized mitochondria and shows reddish-orange fluorescence. The emission intensity ratio of the 545 nm and 595 nm peaks was used as a measure of the mitochondrial depolarization; a higher ratio indicated more depolarization. Hep3B cells with QNT11 treatment for 24 h exhibited green JC-1 fluorescence in a dose-dependent manner, which is consistent with a loss of △ψm (Figure 5). After treatment with QNT11 at 0.803 μM, 2.210 μM and 3.616 μM for 24 h, △ψm of the cells decreased (9.65 ± 4.26)%, (37.34 ± 3.98)% and (45.05 ± 3.18)%, respectively, as compared to the control (P < 0.05,T = 3.24).


Trimethoxy-benzaldehyde levofloxacin hydrazone inducing the growth arrest and apoptosis of human hepatocarcinoma cells.

Sun JP, Shi ZY, Liu SM, Kang YH, Hu GQ, Huangfu CS, Deng JB, Liu B - Cancer Cell Int. (2013)

Effects of QNT11 on the mitochondrial membrane potential of Hep3B cells HCS analysis after JC-1 staining to detect changes in the mitochondrial membrane potential of Hep3B cells Induced by QNT11. Cells were treated with 0, 0.803 (IC30 group), 2.210 (IC50 group)and 3.616 μM (IC70 group) QNT11 respectively for 24 h. The mitochondrial membrane potential (△ψm) was measured by HCS image system.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3716872&req=5

Figure 5: Effects of QNT11 on the mitochondrial membrane potential of Hep3B cells HCS analysis after JC-1 staining to detect changes in the mitochondrial membrane potential of Hep3B cells Induced by QNT11. Cells were treated with 0, 0.803 (IC30 group), 2.210 (IC50 group)and 3.616 μM (IC70 group) QNT11 respectively for 24 h. The mitochondrial membrane potential (△ψm) was measured by HCS image system.
Mentions: Mitochondrial membrane potential (△ψm) was detected with the fluorescent probe JC-1, which exists predominantly in monomeric form in cells with depolarized mitochondria and displays green fluorescence at 490 nm. On the other hand, JC-1 primarily forms aggregates in cells with polarized mitochondria and shows reddish-orange fluorescence. The emission intensity ratio of the 545 nm and 595 nm peaks was used as a measure of the mitochondrial depolarization; a higher ratio indicated more depolarization. Hep3B cells with QNT11 treatment for 24 h exhibited green JC-1 fluorescence in a dose-dependent manner, which is consistent with a loss of △ψm (Figure 5). After treatment with QNT11 at 0.803 μM, 2.210 μM and 3.616 μM for 24 h, △ψm of the cells decreased (9.65 ± 4.26)%, (37.34 ± 3.98)% and (45.05 ± 3.18)%, respectively, as compared to the control (P < 0.05,T = 3.24).

Bottom Line: There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment.In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, 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.The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Nursing, Institute of Neurobiology, Henan University, Kaifeng, China. kangyh0811@gmail.com.

ABSTRACT

Background: In order to search for new structural modification strategies on fluoroquinolones, we have designed and synthesized a series of fluoroquinolone derivatives by linking various hydrazine compounds to the C-3 carboxyl group of levofloxacin and assessed their anticancer activities. Several novel levofloxacin derivatives displayed potent cytotoxicity against the tested cancer cell lines in vitro. In the present study, we investigated the effect of 1-Cyclopropyl-6-fluoro-4-oxo-7- piperazin-1, 4-dihydro- quinoline- 3-carboxylic acid benzo [1,3] dioxol-5- ylmethylene- hydrazide (QNT11) on the apoptosis of human hepatocarcinoma cells in vitro.

Methods: The inhibition effects of QNT11 on cell proliferation were examined by MTT assay. Cell apoptosis was determined by TUNEL and DNA agarose gel electrophoresis method. The topoisomerase ΙΙ activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Cell cycle progression was analyzed using flow cytometry in conjunction with ethanol fixation and propidium iodide staining. Mitochondrial membrane potential (△ψm) was measured by high content screening image system. The caspase-9, caspase-8, caspase-3, Bcl-2, Bax, CDK1, Cyclin B1and cytochrome c protein expressions were detected by Western blot analysis.

Results: QNT11 showed selective cytotoxicity against Hep3B, SMMC-7721, MCF-7 and HCT-8 cell lines with IC50 values of 2.21 μM, 2.38 μM, 3.17 μM and 2.79 μM, respectively. In contrast, QNT11 had weak cytotoxicity against mouse bone marrow mesenchymal stem cells (BMSCs) with IC50 value of 7.46 μM. Treatment of Hep3B cells with different concentrations of QNT11 increased the percentage of the apoptosis cells significantly, and agarose gel electrophoresis revealed the ladder DNA bands typical of apoptotic cells, with a decrease in the mitochondrial membrane potential. Compared to the control group, QNT11 could influence the DNA topoisomerase IIactivity and inhibit the religation of DNA strands, thus keeping the DNA in fragments. There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment. Observed changes in cell cycle distribution by QNT11 treated might be caused by insufficient preparation for G2/M transition. In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, 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.

Conclusions: Our results showed that QNT11 as a fluoroquinolone derivative exerted potent and selectively anticancer activity through the mechanism of eukaryotic topoisomerase II poisoning. The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

No MeSH data available.


Related in: MedlinePlus