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N-acetylcysteine improves antitumoural response of Interferon alpha by NF-kB downregulation in liver cancer cells.

Kretzmann NA, Chiela E, Matte U, Marroni N, Marroni CA - Comp Hepatol (2012)

Bottom Line: More importantly, NAC potentiates the cytotoxic effect of IFN, with the best response achieved with 10 mM of NAC and 2.5 x 104 of IFN.These results were confirmed by Annexin/PI staining through flow cytometry and morphologic analyses.Co-treatment reduced the expression of the nuclear transcription factor kappa-B (NF-kB).

View Article: PubMed Central - HTML - PubMed

Affiliation: Post-Graduation Program in Medicine: Hepatology, Universidade Federal de Ciências da Saúde de Porto Alegre, Brazil, Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre, RS, CEP: 90050-170, Brazil. nakfilho@gmail.com.

ABSTRACT

Background: Liver cancer is one of the most common malignancies in the world and at the moment, there is no drug intervention effective for the treatment of liver tumours. Investigate the effect of N-acetylcysteine (NAC), which has been studied for its antitumoural properties, on the toxicity of hepatocarcinoma (HCC) cells in vitro when used with the drug interferon alpha-2A (IFN), which is used clinically to treat HCC.

Results: NAC, IFN and NAC plus IFN reduced cell viability, as determined by MTT assay. More importantly, NAC potentiates the cytotoxic effect of IFN, with the best response achieved with 10 mM of NAC and 2.5 x 104 of IFN. These results were confirmed by Annexin/PI staining through flow cytometry and morphologic analyses. Co-treatment reduced the expression of the nuclear transcription factor kappa-B (NF-kB). In a similar way to NAC, RNAi against p65 potentiated the toxic effect of IFN, suggesting that, indeed, NAC may be enhancing the effect of IFN through inhibition of NF-kB.

Conclusions: Our results support the notion that NAC may be an important drug for the treatment of liver tumours as primary or adjuvant therapy. IFN has a limited clinical response, and therefore, the anti-proliferative properties of NAC in the liver should be explored further as an alternative for non-responders to IFN treatment.

No MeSH data available.


Related in: MedlinePlus

NAC and IFN treatment induce apoptosis in HCC cells. Cells were treated with IFN 2.5x104 U/mL and/or NAC 10 mM for the indicated time periods. Fluorescence microscopy of HepG2 and Huh7 cells stained with annexin and PI. FITC-labelled Annexin V show apoptotic cells (green) and PI stained cells (red) indicate necrosis or late apoptosis. DAPI stained nuclei (blue). Magnification used was 60x. Bar, 25μm.
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Figure 5: NAC and IFN treatment induce apoptosis in HCC cells. Cells were treated with IFN 2.5x104 U/mL and/or NAC 10 mM for the indicated time periods. Fluorescence microscopy of HepG2 and Huh7 cells stained with annexin and PI. FITC-labelled Annexin V show apoptotic cells (green) and PI stained cells (red) indicate necrosis or late apoptosis. DAPI stained nuclei (blue). Magnification used was 60x. Bar, 25μm.

Mentions: On annexin V/PI analysis through fluorescence microscopy and flow cytometry, both NAC and IFN-α seemed to have proapoptotic effects in both cell lines (Figures5,6 and7). Interestingly, cells presented a different profile of sensitivity to treatments. HepG2 cells were more sensitive to treatment with NAC, presenting positive annexin-V staining at 24 h of treatment, while Huh7 cells were more sensitive to IFN. NAC potentiated the proapoptotic effect of IFN mainly in HepG2 cells, in which the reduction in NF-kB expression was also higher with co-treatment (Figures3 and4).


N-acetylcysteine improves antitumoural response of Interferon alpha by NF-kB downregulation in liver cancer cells.

Kretzmann NA, Chiela E, Matte U, Marroni N, Marroni CA - Comp Hepatol (2012)

NAC and IFN treatment induce apoptosis in HCC cells. Cells were treated with IFN 2.5x104 U/mL and/or NAC 10 mM for the indicated time periods. Fluorescence microscopy of HepG2 and Huh7 cells stained with annexin and PI. FITC-labelled Annexin V show apoptotic cells (green) and PI stained cells (red) indicate necrosis or late apoptosis. DAPI stained nuclei (blue). Magnification used was 60x. Bar, 25μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3539937&req=5

Figure 5: NAC and IFN treatment induce apoptosis in HCC cells. Cells were treated with IFN 2.5x104 U/mL and/or NAC 10 mM for the indicated time periods. Fluorescence microscopy of HepG2 and Huh7 cells stained with annexin and PI. FITC-labelled Annexin V show apoptotic cells (green) and PI stained cells (red) indicate necrosis or late apoptosis. DAPI stained nuclei (blue). Magnification used was 60x. Bar, 25μm.
Mentions: On annexin V/PI analysis through fluorescence microscopy and flow cytometry, both NAC and IFN-α seemed to have proapoptotic effects in both cell lines (Figures5,6 and7). Interestingly, cells presented a different profile of sensitivity to treatments. HepG2 cells were more sensitive to treatment with NAC, presenting positive annexin-V staining at 24 h of treatment, while Huh7 cells were more sensitive to IFN. NAC potentiated the proapoptotic effect of IFN mainly in HepG2 cells, in which the reduction in NF-kB expression was also higher with co-treatment (Figures3 and4).

Bottom Line: More importantly, NAC potentiates the cytotoxic effect of IFN, with the best response achieved with 10 mM of NAC and 2.5 x 104 of IFN.These results were confirmed by Annexin/PI staining through flow cytometry and morphologic analyses.Co-treatment reduced the expression of the nuclear transcription factor kappa-B (NF-kB).

View Article: PubMed Central - HTML - PubMed

Affiliation: Post-Graduation Program in Medicine: Hepatology, Universidade Federal de Ciências da Saúde de Porto Alegre, Brazil, Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre, RS, CEP: 90050-170, Brazil. nakfilho@gmail.com.

ABSTRACT

Background: Liver cancer is one of the most common malignancies in the world and at the moment, there is no drug intervention effective for the treatment of liver tumours. Investigate the effect of N-acetylcysteine (NAC), which has been studied for its antitumoural properties, on the toxicity of hepatocarcinoma (HCC) cells in vitro when used with the drug interferon alpha-2A (IFN), which is used clinically to treat HCC.

Results: NAC, IFN and NAC plus IFN reduced cell viability, as determined by MTT assay. More importantly, NAC potentiates the cytotoxic effect of IFN, with the best response achieved with 10 mM of NAC and 2.5 x 104 of IFN. These results were confirmed by Annexin/PI staining through flow cytometry and morphologic analyses. Co-treatment reduced the expression of the nuclear transcription factor kappa-B (NF-kB). In a similar way to NAC, RNAi against p65 potentiated the toxic effect of IFN, suggesting that, indeed, NAC may be enhancing the effect of IFN through inhibition of NF-kB.

Conclusions: Our results support the notion that NAC may be an important drug for the treatment of liver tumours as primary or adjuvant therapy. IFN has a limited clinical response, and therefore, the anti-proliferative properties of NAC in the liver should be explored further as an alternative for non-responders to IFN treatment.

No MeSH data available.


Related in: MedlinePlus