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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

Knock down of p65 subunit shown by real-time PCR. Relative quantification of p65 normalised by the expression of GAPDH in HepG2 and Huh7 cells 24 hours after transfection. Values are shown as means and standard errors of the mean (SEM). a- siRNAp65x COsiRNA p<0.01-HepG2. b- siRNAp65x COsiRNA p<0.01-Huh7.
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Figure 8: Knock down of p65 subunit shown by real-time PCR. Relative quantification of p65 normalised by the expression of GAPDH in HepG2 and Huh7 cells 24 hours after transfection. Values are shown as means and standard errors of the mean (SEM). a- siRNAp65x COsiRNA p<0.01-HepG2. b- siRNAp65x COsiRNA p<0.01-Huh7.

Mentions: We then explored the role of the NF-kB pathway on NAC and IFN-α toxicity using siRNA-mediated p65 knockdown (KD cells). At 24 h post-transfection, a greater reduction of 95% of p65 expression levels was observed both through fluorescence microscopy (data not shown) and real-time PCR (Figure8).


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)

Knock down of p65 subunit shown by real-time PCR. Relative quantification of p65 normalised by the expression of GAPDH in HepG2 and Huh7 cells 24 hours after transfection. Values are shown as means and standard errors of the mean (SEM). a- siRNAp65x COsiRNA p<0.01-HepG2. b- siRNAp65x COsiRNA p<0.01-Huh7.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 8: Knock down of p65 subunit shown by real-time PCR. Relative quantification of p65 normalised by the expression of GAPDH in HepG2 and Huh7 cells 24 hours after transfection. Values are shown as means and standard errors of the mean (SEM). a- siRNAp65x COsiRNA p<0.01-HepG2. b- siRNAp65x COsiRNA p<0.01-Huh7.
Mentions: We then explored the role of the NF-kB pathway on NAC and IFN-α toxicity using siRNA-mediated p65 knockdown (KD cells). At 24 h post-transfection, a greater reduction of 95% of p65 expression levels was observed both through fluorescence microscopy (data not shown) and real-time PCR (Figure8).

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