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Evaluation of the antioxidant activity and antiproliferative effect of the jaboticaba (Myrciaria cauliflora) seed extracts in oral carcinoma cells.

Wang WH, Tyan YC, Chen ZS, Lin CG, Yang MH, Yuan SS, Tsai WC - Biomed Res Int (2014)

Bottom Line: Water extracts of jaboticaba seeds showed concentration-dependent antiproliferative effects.The antioxidant activity of jaboticaba was investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays, and the drug concentration eliciting 50% maximum stimulation (SC50) was determined.The present findings suggest that water extracts of jaboticaba seeds exhibit an antiproliferative effect against oral cancer cells by inducing apoptosis through downregulating survivin expression and thereby activating caspase-mediated Bid cleavage.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Cathay General Hospital, Taipei City 106, Taiwan ; Department of Otolaryngology, Sijhih Cathay General Hospital, New Taipei City 221, Taiwan ; School of Medicine, Fu-Jen Catholic University, New Taipei City 242, Taiwan.

ABSTRACT
It is becoming increasingly evident that certain phytochemicals possess cancer chemopreventive properties. In this study, the antiproliferative activity of extracts from different parts of the jaboticaba (Myrciaria cauliflora) plant was evaluated for its effect on human oral carcinoma cell lines. The cytotoxicities of various plant extract concentrations were examined and the 50% maximal inhibitory concentration (IC50) was determined. Water extracts of jaboticaba seeds showed concentration-dependent antiproliferative effects. Annexin V/propidium iodide positivity with active caspase-3 induction indicated that the treated cells underwent apoptosis. Several important regulatory proteins (Bcl-2, Bcl-xL, Bid, and survivin) involved in apoptosis were also evaluated. The antioxidant activity of jaboticaba was investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays, and the drug concentration eliciting 50% maximum stimulation (SC50) was determined. The present findings suggest that water extracts of jaboticaba seeds exhibit an antiproliferative effect against oral cancer cells by inducing apoptosis through downregulating survivin expression and thereby activating caspase-mediated Bid cleavage.

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Related in: MedlinePlus

Effects of jaboticaba seed water extract on apoptosis regulatory proteins in HSC-3 cells. HSC-3 cells were treated with 5, 10, 25, 50, and 100 μg/ml of jaboticaba seed extract or ddH2O as a control for 48 h, and then total proteins were isolated. Equal amounts of cell lysates were analyzed for PARP, survivin, Bid, Bcl-xL, and Bcl-2 expression by Western blotting with corresponding antibodies. β-actin served as the loading control. The fold change was calculated as the ratio of the target proteins in the presence of indicated concentration of Jaboticaba seed extract after normalization of the target proteins to β-actin in each lane.
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fig4: Effects of jaboticaba seed water extract on apoptosis regulatory proteins in HSC-3 cells. HSC-3 cells were treated with 5, 10, 25, 50, and 100 μg/ml of jaboticaba seed extract or ddH2O as a control for 48 h, and then total proteins were isolated. Equal amounts of cell lysates were analyzed for PARP, survivin, Bid, Bcl-xL, and Bcl-2 expression by Western blotting with corresponding antibodies. β-actin served as the loading control. The fold change was calculated as the ratio of the target proteins in the presence of indicated concentration of Jaboticaba seed extract after normalization of the target proteins to β-actin in each lane.

Mentions: Treatment of HSC-3 cells with different concentrations of water extracts of jaboticaba seeds for 24 h promoted dose-dependent cleavage of poly (ADP-ribose) polymerase (PARP) from the full length 116-kDa to an inactive 85-kDa form by activating caspases (Figure 4), which is another indicator of apoptosis. In order to further understand the apoptotic phenomenon, we evaluated the protein level of various key regulators in the apoptosis pathway through Western blot analysis. It is known that caspase-3 activity can be inhibited by a group of proteins that are collectively termed “inhibitors of apoptosis proteins,” of which survivin is one. We particularly determined the expression of survivin because it was shown to directly bind and inhibit caspase-3 [14, 15]. The dramatic abolishment of survivin thereby activated Bid cleavage, indicating that the water extract of jaboticaba seeds induced cell death by lowering the inhibition of apoptosis. Remarkably, the conventional intrinsic apoptosis pathway controlled by the Bcl-2 family did not show an unbalanced change.


Evaluation of the antioxidant activity and antiproliferative effect of the jaboticaba (Myrciaria cauliflora) seed extracts in oral carcinoma cells.

Wang WH, Tyan YC, Chen ZS, Lin CG, Yang MH, Yuan SS, Tsai WC - Biomed Res Int (2014)

Effects of jaboticaba seed water extract on apoptosis regulatory proteins in HSC-3 cells. HSC-3 cells were treated with 5, 10, 25, 50, and 100 μg/ml of jaboticaba seed extract or ddH2O as a control for 48 h, and then total proteins were isolated. Equal amounts of cell lysates were analyzed for PARP, survivin, Bid, Bcl-xL, and Bcl-2 expression by Western blotting with corresponding antibodies. β-actin served as the loading control. The fold change was calculated as the ratio of the target proteins in the presence of indicated concentration of Jaboticaba seed extract after normalization of the target proteins to β-actin in each lane.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150497&req=5

fig4: Effects of jaboticaba seed water extract on apoptosis regulatory proteins in HSC-3 cells. HSC-3 cells were treated with 5, 10, 25, 50, and 100 μg/ml of jaboticaba seed extract or ddH2O as a control for 48 h, and then total proteins were isolated. Equal amounts of cell lysates were analyzed for PARP, survivin, Bid, Bcl-xL, and Bcl-2 expression by Western blotting with corresponding antibodies. β-actin served as the loading control. The fold change was calculated as the ratio of the target proteins in the presence of indicated concentration of Jaboticaba seed extract after normalization of the target proteins to β-actin in each lane.
Mentions: Treatment of HSC-3 cells with different concentrations of water extracts of jaboticaba seeds for 24 h promoted dose-dependent cleavage of poly (ADP-ribose) polymerase (PARP) from the full length 116-kDa to an inactive 85-kDa form by activating caspases (Figure 4), which is another indicator of apoptosis. In order to further understand the apoptotic phenomenon, we evaluated the protein level of various key regulators in the apoptosis pathway through Western blot analysis. It is known that caspase-3 activity can be inhibited by a group of proteins that are collectively termed “inhibitors of apoptosis proteins,” of which survivin is one. We particularly determined the expression of survivin because it was shown to directly bind and inhibit caspase-3 [14, 15]. The dramatic abolishment of survivin thereby activated Bid cleavage, indicating that the water extract of jaboticaba seeds induced cell death by lowering the inhibition of apoptosis. Remarkably, the conventional intrinsic apoptosis pathway controlled by the Bcl-2 family did not show an unbalanced change.

Bottom Line: Water extracts of jaboticaba seeds showed concentration-dependent antiproliferative effects.The antioxidant activity of jaboticaba was investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays, and the drug concentration eliciting 50% maximum stimulation (SC50) was determined.The present findings suggest that water extracts of jaboticaba seeds exhibit an antiproliferative effect against oral cancer cells by inducing apoptosis through downregulating survivin expression and thereby activating caspase-mediated Bid cleavage.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, Cathay General Hospital, Taipei City 106, Taiwan ; Department of Otolaryngology, Sijhih Cathay General Hospital, New Taipei City 221, Taiwan ; School of Medicine, Fu-Jen Catholic University, New Taipei City 242, Taiwan.

ABSTRACT
It is becoming increasingly evident that certain phytochemicals possess cancer chemopreventive properties. In this study, the antiproliferative activity of extracts from different parts of the jaboticaba (Myrciaria cauliflora) plant was evaluated for its effect on human oral carcinoma cell lines. The cytotoxicities of various plant extract concentrations were examined and the 50% maximal inhibitory concentration (IC50) was determined. Water extracts of jaboticaba seeds showed concentration-dependent antiproliferative effects. Annexin V/propidium iodide positivity with active caspase-3 induction indicated that the treated cells underwent apoptosis. Several important regulatory proteins (Bcl-2, Bcl-xL, Bid, and survivin) involved in apoptosis were also evaluated. The antioxidant activity of jaboticaba was investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays, and the drug concentration eliciting 50% maximum stimulation (SC50) was determined. The present findings suggest that water extracts of jaboticaba seeds exhibit an antiproliferative effect against oral cancer cells by inducing apoptosis through downregulating survivin expression and thereby activating caspase-mediated Bid cleavage.

Show MeSH
Related in: MedlinePlus