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Lipid droplet binding thalidomide analogs activate endoplasmic reticulum stress and suppress hepatocellular carcinoma in a chemically induced transgenic mouse model.

Nagy LI, Molnár E, Kanizsai I, Madácsi R, Ózsvári B, Fehér LZ, Fábián G, Marton A, Vizler C, Ayaydin F, Kitajka K, Hackler L, Mátés L, Deák F, Kiss I, Puskás LG - Lipids Health Dis (2013)

Bottom Line: In this study, we report the in vitro and in vivo effects of two novel amino-trifluoro-phtalimide analogs, Ac-915 and Ac-2010.The analogs mainly accumulated in the liver.These results imply that these amino-trifluoro-phthalimide analogs could serve potent clinical candidates against HCC alone or in combination with dietary polyunsaturated fatty acids.

View Article: PubMed Central - HTML - PubMed

Affiliation: AVIDIN Ltd,, Szeged, Hungary. puskas.szbk@gmail.com.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is the most frequent and aggressive primary tumor of the liver and it has limited treatment options.

Results: In this study, we report the in vitro and in vivo effects of two novel amino-trifluoro-phtalimide analogs, Ac-915 and Ac-2010. Both compounds bind lipid droplets and endoplasmic reticulum membrane, and interact with several proteins with chaperone functions (HSP60, HSP70, HSP90, and protein disulfide isomerase) as determined by affinity chromatography and resonant waveguide optical biosensor technology. Both compounds inhibited protein disulfide isomerase activity and induced cell death of different HCC cells at sub or low micromolar ranges detected by classical biochemical end-point assay as well as with real-time label-free measurements. Besides cell proliferation inhibiton, analogs also inhibited cell migration even at 250 nM. Relative biodistribution of the analogs was analysed in native tissue sections of different organs after administration of drugs, and by using fluorescent confocal microscopy based on the inherent blue fluorescence of the compounds. The analogs mainly accumulated in the liver. The effects of Ac-915 and Ac-2010 were also demonstrated on the advanced stages of hepatocarcinogenesis in a transgenic mouse model of N-nitrosodiethylamine (DEN)-induced HCC. Significantly less tumor development was found in the livers of the Ac-915- or Ac-2010-treated groups compared with control mice, characterized by less liver tumor incidence, fewer tumors and smaller tumor size.

Conclusion: These results imply that these amino-trifluoro-phthalimide analogs could serve potent clinical candidates against HCC alone or in combination with dietary polyunsaturated fatty acids.

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Related in: MedlinePlus

Intracellular glutathione (GSH) concentrations in Ac-915 treated Hep3B cells. (a) GSH content is shown relative to values obtained with the untreated control (%). (b) ROS formation (DCFDA %) and cell death (PI %) after treatment of Hep3B cells with Ac-915 at different concentrations 30 min and 60 min after treatment.
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Figure 4: Intracellular glutathione (GSH) concentrations in Ac-915 treated Hep3B cells. (a) GSH content is shown relative to values obtained with the untreated control (%). (b) ROS formation (DCFDA %) and cell death (PI %) after treatment of Hep3B cells with Ac-915 at different concentrations 30 min and 60 min after treatment.

Mentions: Toxic compounds and numerous anticancer agents interfere with chaperone and ER functions leading to cellular stress which is manifested by elevated reactive oxygen species (ROS) and dramatic decrease in the anti-oxidant, glutathione (GSH) level. To investigate whether Ac-915 and Ac-2010 exert a pro-oxidative effect as determined earlier for other amino-trifluoro-phthalimides [4] and “redox-reactive” thalidomide analogs [27], we measured the presence of ROS in human hepatocellular carcinoma cells (Hep3B) Figure 4a. To reveal the correlation of depletion of glutathione and ROS production of the analogs we determined the intracellular concentrations of glutathione. To determine whether Ac-915 and Ac-2010 affect intracellular GSH levels Hep3B cells were treated with compounds and GSH levels were recorded. According to our expectations, by inducing oxidative stress both compounds also depleted intracellular GSH levels (for Ac-915 GSH levels, see Figure 4b.


Lipid droplet binding thalidomide analogs activate endoplasmic reticulum stress and suppress hepatocellular carcinoma in a chemically induced transgenic mouse model.

Nagy LI, Molnár E, Kanizsai I, Madácsi R, Ózsvári B, Fehér LZ, Fábián G, Marton A, Vizler C, Ayaydin F, Kitajka K, Hackler L, Mátés L, Deák F, Kiss I, Puskás LG - Lipids Health Dis (2013)

Intracellular glutathione (GSH) concentrations in Ac-915 treated Hep3B cells. (a) GSH content is shown relative to values obtained with the untreated control (%). (b) ROS formation (DCFDA %) and cell death (PI %) after treatment of Hep3B cells with Ac-915 at different concentrations 30 min and 60 min after treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4222488&req=5

Figure 4: Intracellular glutathione (GSH) concentrations in Ac-915 treated Hep3B cells. (a) GSH content is shown relative to values obtained with the untreated control (%). (b) ROS formation (DCFDA %) and cell death (PI %) after treatment of Hep3B cells with Ac-915 at different concentrations 30 min and 60 min after treatment.
Mentions: Toxic compounds and numerous anticancer agents interfere with chaperone and ER functions leading to cellular stress which is manifested by elevated reactive oxygen species (ROS) and dramatic decrease in the anti-oxidant, glutathione (GSH) level. To investigate whether Ac-915 and Ac-2010 exert a pro-oxidative effect as determined earlier for other amino-trifluoro-phthalimides [4] and “redox-reactive” thalidomide analogs [27], we measured the presence of ROS in human hepatocellular carcinoma cells (Hep3B) Figure 4a. To reveal the correlation of depletion of glutathione and ROS production of the analogs we determined the intracellular concentrations of glutathione. To determine whether Ac-915 and Ac-2010 affect intracellular GSH levels Hep3B cells were treated with compounds and GSH levels were recorded. According to our expectations, by inducing oxidative stress both compounds also depleted intracellular GSH levels (for Ac-915 GSH levels, see Figure 4b.

Bottom Line: In this study, we report the in vitro and in vivo effects of two novel amino-trifluoro-phtalimide analogs, Ac-915 and Ac-2010.The analogs mainly accumulated in the liver.These results imply that these amino-trifluoro-phthalimide analogs could serve potent clinical candidates against HCC alone or in combination with dietary polyunsaturated fatty acids.

View Article: PubMed Central - HTML - PubMed

Affiliation: AVIDIN Ltd,, Szeged, Hungary. puskas.szbk@gmail.com.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is the most frequent and aggressive primary tumor of the liver and it has limited treatment options.

Results: In this study, we report the in vitro and in vivo effects of two novel amino-trifluoro-phtalimide analogs, Ac-915 and Ac-2010. Both compounds bind lipid droplets and endoplasmic reticulum membrane, and interact with several proteins with chaperone functions (HSP60, HSP70, HSP90, and protein disulfide isomerase) as determined by affinity chromatography and resonant waveguide optical biosensor technology. Both compounds inhibited protein disulfide isomerase activity and induced cell death of different HCC cells at sub or low micromolar ranges detected by classical biochemical end-point assay as well as with real-time label-free measurements. Besides cell proliferation inhibiton, analogs also inhibited cell migration even at 250 nM. Relative biodistribution of the analogs was analysed in native tissue sections of different organs after administration of drugs, and by using fluorescent confocal microscopy based on the inherent blue fluorescence of the compounds. The analogs mainly accumulated in the liver. The effects of Ac-915 and Ac-2010 were also demonstrated on the advanced stages of hepatocarcinogenesis in a transgenic mouse model of N-nitrosodiethylamine (DEN)-induced HCC. Significantly less tumor development was found in the livers of the Ac-915- or Ac-2010-treated groups compared with control mice, characterized by less liver tumor incidence, fewer tumors and smaller tumor size.

Conclusion: These results imply that these amino-trifluoro-phthalimide analogs could serve potent clinical candidates against HCC alone or in combination with dietary polyunsaturated fatty acids.

Show MeSH
Related in: MedlinePlus