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Trans-1,3-diphenyl-2,3-epoxypropan-1-one, a chalcone derivative, induces apoptosis via ROS-mediated down-regulation of Bcl-xL in human leukemia HL-60 cells.

Ko EY, Lee SH, Ko JY, Moon JY, Yoon WJ, Ahn G, Roh SW, Cho K, Jeon YJ, Kim D, Kim KN - EXCLI J (2015)

Bottom Line: Treatment of HL-60 cells with various concentration of DPEP resulted in a sequence of events characteristic of apoptosis, including loss of cell viability, morphological changes, and increased sub-G1 DNA content.However, NAC pre-treatment significantly inhibited the activation of caspase-3 and PARP cleavage and reduced Bcl-xL levels.These findings provide the first evidence that DPEP may inhibit the growth of HL-60 cells and induce apoptosis through a ROS-mediated Bcl-xL pathway.

View Article: PubMed Central - PubMed

Affiliation: Jeju Center, Korea Basic Science Institute (KBSI), Jeju 690-140, Republic of Korea; School of Marine Biomedical Sciences, Jeju National University, Jeju 690-756, Republic of Korea.

ABSTRACT
The anticancer effects of trans-1,3-diphenyl-2,3-epoxypropan-1-one (DPEP), a chalcone derivative, were investigated in human leukemia HL-60 cells. Treatment of HL-60 cells with various concentration of DPEP resulted in a sequence of events characteristic of apoptosis, including loss of cell viability, morphological changes, and increased sub-G1 DNA content. We demonstrated that DPEP elevates reactive oxygen species (ROS) levels in HL-60 cells, and that the ROS scavenger N-acetylcysteine (NAC) could block DPEP-induced ROS generation and apoptosis. Western blot analysis revealed that DPEP inhibits Bcl-xL expression, leading to caspase-3 activation and poly-ADP-ribose polymerase (PARP) cleavage, thereby inducing apoptosis. However, NAC pre-treatment significantly inhibited the activation of caspase-3 and PARP cleavage and reduced Bcl-xL levels. These findings provide the first evidence that DPEP may inhibit the growth of HL-60 cells and induce apoptosis through a ROS-mediated Bcl-xL pathway.

No MeSH data available.


Related in: MedlinePlus

NAC pretreatment protected against DPEP-induced apoptosis via the suppression of ROS generation. HL-60 cells were pretreated with 1 mM NAC for 1 h prior to 2 h (A), 12 h (B, C), or 24 h (D) of 40 μM DPEP exposure. (A) The cells were labeled with 10 μM DCFH2-DA for 30 min at 37 °C, and subjected to subsequent FACS analyses for intracellular ROS accumulation. (B) Apoptotic bodies were stained with Hoechst 33342 solution and observed under a fluorescent microscope using a blue filter. (C) The cells were stained with PI and analyzed via flow cytometry. (D) Cell viability was measured via MTT assay. Each value indicates the mean ± standard error from three independent experiments. Difference with *p < 0.05 is considered statistically significant.
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Figure 3: NAC pretreatment protected against DPEP-induced apoptosis via the suppression of ROS generation. HL-60 cells were pretreated with 1 mM NAC for 1 h prior to 2 h (A), 12 h (B, C), or 24 h (D) of 40 μM DPEP exposure. (A) The cells were labeled with 10 μM DCFH2-DA for 30 min at 37 °C, and subjected to subsequent FACS analyses for intracellular ROS accumulation. (B) Apoptotic bodies were stained with Hoechst 33342 solution and observed under a fluorescent microscope using a blue filter. (C) The cells were stained with PI and analyzed via flow cytometry. (D) Cell viability was measured via MTT assay. Each value indicates the mean ± standard error from three independent experiments. Difference with *p < 0.05 is considered statistically significant.

Mentions: We used the fluorescent probe DCFH-DA to measure the generation of intracellular ROS in DPEP-treated HL-60 cells. As shown in Figure 3A(Fig. 3), DPEP significantly increased ROS production, as indicated by a rightward shift in fluorescence during the flow cytometric analysis. Moreover, pretreatment with the antioxidant NAC significantly reduced intracellular ROS levels compared to cells treated with DPEP alone. The inhibition of ROS generation by NAC also markedly prevented DPEP-induced apoptotic bodies (Figure 3B(Fig. 3)) and increased the percentage of sub-G1 cells (Figure 3C(Fig. 3)). NAC also inhibited DPEP-induced reductions in cell viability in HL-60 cells (Figure 3D(Fig. 3)). Together, these results suggest that intracellular ROS play an essential role in cell death by DPEP-induced apoptosis in HL-60 cells.


Trans-1,3-diphenyl-2,3-epoxypropan-1-one, a chalcone derivative, induces apoptosis via ROS-mediated down-regulation of Bcl-xL in human leukemia HL-60 cells.

Ko EY, Lee SH, Ko JY, Moon JY, Yoon WJ, Ahn G, Roh SW, Cho K, Jeon YJ, Kim D, Kim KN - EXCLI J (2015)

NAC pretreatment protected against DPEP-induced apoptosis via the suppression of ROS generation. HL-60 cells were pretreated with 1 mM NAC for 1 h prior to 2 h (A), 12 h (B, C), or 24 h (D) of 40 μM DPEP exposure. (A) The cells were labeled with 10 μM DCFH2-DA for 30 min at 37 °C, and subjected to subsequent FACS analyses for intracellular ROS accumulation. (B) Apoptotic bodies were stained with Hoechst 33342 solution and observed under a fluorescent microscope using a blue filter. (C) The cells were stained with PI and analyzed via flow cytometry. (D) Cell viability was measured via MTT assay. Each value indicates the mean ± standard error from three independent experiments. Difference with *p < 0.05 is considered statistically significant.
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Related In: Results  -  Collection

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Figure 3: NAC pretreatment protected against DPEP-induced apoptosis via the suppression of ROS generation. HL-60 cells were pretreated with 1 mM NAC for 1 h prior to 2 h (A), 12 h (B, C), or 24 h (D) of 40 μM DPEP exposure. (A) The cells were labeled with 10 μM DCFH2-DA for 30 min at 37 °C, and subjected to subsequent FACS analyses for intracellular ROS accumulation. (B) Apoptotic bodies were stained with Hoechst 33342 solution and observed under a fluorescent microscope using a blue filter. (C) The cells were stained with PI and analyzed via flow cytometry. (D) Cell viability was measured via MTT assay. Each value indicates the mean ± standard error from three independent experiments. Difference with *p < 0.05 is considered statistically significant.
Mentions: We used the fluorescent probe DCFH-DA to measure the generation of intracellular ROS in DPEP-treated HL-60 cells. As shown in Figure 3A(Fig. 3), DPEP significantly increased ROS production, as indicated by a rightward shift in fluorescence during the flow cytometric analysis. Moreover, pretreatment with the antioxidant NAC significantly reduced intracellular ROS levels compared to cells treated with DPEP alone. The inhibition of ROS generation by NAC also markedly prevented DPEP-induced apoptotic bodies (Figure 3B(Fig. 3)) and increased the percentage of sub-G1 cells (Figure 3C(Fig. 3)). NAC also inhibited DPEP-induced reductions in cell viability in HL-60 cells (Figure 3D(Fig. 3)). Together, these results suggest that intracellular ROS play an essential role in cell death by DPEP-induced apoptosis in HL-60 cells.

Bottom Line: Treatment of HL-60 cells with various concentration of DPEP resulted in a sequence of events characteristic of apoptosis, including loss of cell viability, morphological changes, and increased sub-G1 DNA content.However, NAC pre-treatment significantly inhibited the activation of caspase-3 and PARP cleavage and reduced Bcl-xL levels.These findings provide the first evidence that DPEP may inhibit the growth of HL-60 cells and induce apoptosis through a ROS-mediated Bcl-xL pathway.

View Article: PubMed Central - PubMed

Affiliation: Jeju Center, Korea Basic Science Institute (KBSI), Jeju 690-140, Republic of Korea; School of Marine Biomedical Sciences, Jeju National University, Jeju 690-756, Republic of Korea.

ABSTRACT
The anticancer effects of trans-1,3-diphenyl-2,3-epoxypropan-1-one (DPEP), a chalcone derivative, were investigated in human leukemia HL-60 cells. Treatment of HL-60 cells with various concentration of DPEP resulted in a sequence of events characteristic of apoptosis, including loss of cell viability, morphological changes, and increased sub-G1 DNA content. We demonstrated that DPEP elevates reactive oxygen species (ROS) levels in HL-60 cells, and that the ROS scavenger N-acetylcysteine (NAC) could block DPEP-induced ROS generation and apoptosis. Western blot analysis revealed that DPEP inhibits Bcl-xL expression, leading to caspase-3 activation and poly-ADP-ribose polymerase (PARP) cleavage, thereby inducing apoptosis. However, NAC pre-treatment significantly inhibited the activation of caspase-3 and PARP cleavage and reduced Bcl-xL levels. These findings provide the first evidence that DPEP may inhibit the growth of HL-60 cells and induce apoptosis through a ROS-mediated Bcl-xL pathway.

No MeSH data available.


Related in: MedlinePlus