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Free Radical Scavenging Activity: Antiproliferative and Proteomics Analyses of the Differential Expression of Apoptotic Proteins in MCF-7 Cells Treated with Acetone Leaf Extract of Diospyros lycioides (Ebenaceae).

Pilane MC, Bagla VP, Mokgotho MP, Mbazima V, Matsebatlela TM, Ncube I, Mampuru L - Evid Based Complement Alternat Med (2015)

Bottom Line: The acetone leaf extract of Diospyros lycioides was evaluated qualitatively and quantitatively for its antioxidant potential using DPPH assay and nitric oxide radical scavenging effect, while the viability of MCF-7 cells was evaluated using the MTT.Cytotoxicity was demonstrated to be due to apoptosis, with 70% of the extract-treated cells being annexin-V-positive/PI negative at 48 hours.The extract was also shown to upregulate the expression of p53 gene with concomitant downregulation of the Bcl-2 antiapoptotic gene while differentially expressed proteins were identified as enolase, pyruvate kinase, and glyceraldehyde-3-phosphate.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Microbiology and Biotechnology, Faculty of Science and Agriculture, University of Limpopo, Turfloop Campus, Private Bag X1106, Sovenga, Limpopo 0727, South Africa.

ABSTRACT
Breast cancer is the most common cancer in South Africa. The acetone leaf extract of Diospyros lycioides was evaluated qualitatively and quantitatively for its antioxidant potential using DPPH assay and nitric oxide radical scavenging effect, while the viability of MCF-7 cells was evaluated using the MTT. MCF-7 treated cells were stained with Hoechst 335258 dye and annexin-V-FITC to be evaluated for apoptotic effect of the extract, while mRNA expression levels of apoptotic genes were assessed by quantitative real-time PCR and deferential protein expression levels using 2D gel electrophoresis and mass spectrometry. Results revealed presence of antioxidant constituents in the extract. Extract was shown to be cytotoxic in a concentration- and time-dependent manner. Cytotoxicity was demonstrated to be due to apoptosis, with 70% of the extract-treated cells being annexin-V-positive/PI negative at 48 hours. The extract was also shown to upregulate the expression of p53 gene with concomitant downregulation of the Bcl-2 antiapoptotic gene while differentially expressed proteins were identified as enolase, pyruvate kinase, and glyceraldehyde-3-phosphate. The extract in this study was shown to induce apoptosis at an early stage which makes it an ideal source that can be explored for compounds that may be used in the treatment and management of cancer.

No MeSH data available.


Related in: MedlinePlus

The nitric oxide free radical scavenging effect of the crude extract of DL. Curcumin (20 μM = 7.36 μg/mL) was used as a positive control.  ∗∗Statistically significant (p < 0.01).
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fig3: The nitric oxide free radical scavenging effect of the crude extract of DL. Curcumin (20 μM = 7.36 μg/mL) was used as a positive control.  ∗∗Statistically significant (p < 0.01).

Mentions: The free radical scavenging activities of the extract were assayed using quantitative DPPH and nitric oxide scavenging activity assays. It appears that the acetone leaf extract of DL possesses constituents with hydrogen donating abilities, indicative of their antioxidant activity in a concentration-dependent manner (Figure 2). An increase in concentration is shown to increase the scavenging activity of the extract in the DPPH radical scavenging assay whereas the reverse was the case in the nitric oxide free radical scavenging assay as measured by Griess reagent (Figure 3). High accumulation of nitric oxide in tissues causes pathologies such as hypertension, cardiovascular dysfunctions, neurodegenerative diseases, arthritis, asthma, and septic shock in human.


Free Radical Scavenging Activity: Antiproliferative and Proteomics Analyses of the Differential Expression of Apoptotic Proteins in MCF-7 Cells Treated with Acetone Leaf Extract of Diospyros lycioides (Ebenaceae).

Pilane MC, Bagla VP, Mokgotho MP, Mbazima V, Matsebatlela TM, Ncube I, Mampuru L - Evid Based Complement Alternat Med (2015)

The nitric oxide free radical scavenging effect of the crude extract of DL. Curcumin (20 μM = 7.36 μg/mL) was used as a positive control.  ∗∗Statistically significant (p < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4589632&req=5

fig3: The nitric oxide free radical scavenging effect of the crude extract of DL. Curcumin (20 μM = 7.36 μg/mL) was used as a positive control.  ∗∗Statistically significant (p < 0.01).
Mentions: The free radical scavenging activities of the extract were assayed using quantitative DPPH and nitric oxide scavenging activity assays. It appears that the acetone leaf extract of DL possesses constituents with hydrogen donating abilities, indicative of their antioxidant activity in a concentration-dependent manner (Figure 2). An increase in concentration is shown to increase the scavenging activity of the extract in the DPPH radical scavenging assay whereas the reverse was the case in the nitric oxide free radical scavenging assay as measured by Griess reagent (Figure 3). High accumulation of nitric oxide in tissues causes pathologies such as hypertension, cardiovascular dysfunctions, neurodegenerative diseases, arthritis, asthma, and septic shock in human.

Bottom Line: The acetone leaf extract of Diospyros lycioides was evaluated qualitatively and quantitatively for its antioxidant potential using DPPH assay and nitric oxide radical scavenging effect, while the viability of MCF-7 cells was evaluated using the MTT.Cytotoxicity was demonstrated to be due to apoptosis, with 70% of the extract-treated cells being annexin-V-positive/PI negative at 48 hours.The extract was also shown to upregulate the expression of p53 gene with concomitant downregulation of the Bcl-2 antiapoptotic gene while differentially expressed proteins were identified as enolase, pyruvate kinase, and glyceraldehyde-3-phosphate.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Microbiology and Biotechnology, Faculty of Science and Agriculture, University of Limpopo, Turfloop Campus, Private Bag X1106, Sovenga, Limpopo 0727, South Africa.

ABSTRACT
Breast cancer is the most common cancer in South Africa. The acetone leaf extract of Diospyros lycioides was evaluated qualitatively and quantitatively for its antioxidant potential using DPPH assay and nitric oxide radical scavenging effect, while the viability of MCF-7 cells was evaluated using the MTT. MCF-7 treated cells were stained with Hoechst 335258 dye and annexin-V-FITC to be evaluated for apoptotic effect of the extract, while mRNA expression levels of apoptotic genes were assessed by quantitative real-time PCR and deferential protein expression levels using 2D gel electrophoresis and mass spectrometry. Results revealed presence of antioxidant constituents in the extract. Extract was shown to be cytotoxic in a concentration- and time-dependent manner. Cytotoxicity was demonstrated to be due to apoptosis, with 70% of the extract-treated cells being annexin-V-positive/PI negative at 48 hours. The extract was also shown to upregulate the expression of p53 gene with concomitant downregulation of the Bcl-2 antiapoptotic gene while differentially expressed proteins were identified as enolase, pyruvate kinase, and glyceraldehyde-3-phosphate. The extract in this study was shown to induce apoptosis at an early stage which makes it an ideal source that can be explored for compounds that may be used in the treatment and management of cancer.

No MeSH data available.


Related in: MedlinePlus