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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

(A) Effect of DPEP on apoptosis-related protein in HL-60 cells. The cells were exposed to 40 μM DPEP for 1-12 h. (B) Effect of NAC on down-regulation of Bcl-xL and caspase-3, and PARP cleavage activation by DPEP. The cells were pretreated with 1 mM NAC and then treated for 8 h with 40 μM DPEP. Whole cell lysates were subjected to Western blot analysis of anti-Bax, -Bcl-xL, -cleaved-caspase -3, and -PARP monoclonal antibodies. β-Actin was used as internal control. The experiment was repeated three independent times.
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Figure 4: (A) Effect of DPEP on apoptosis-related protein in HL-60 cells. The cells were exposed to 40 μM DPEP for 1-12 h. (B) Effect of NAC on down-regulation of Bcl-xL and caspase-3, and PARP cleavage activation by DPEP. The cells were pretreated with 1 mM NAC and then treated for 8 h with 40 μM DPEP. Whole cell lysates were subjected to Western blot analysis of anti-Bax, -Bcl-xL, -cleaved-caspase -3, and -PARP monoclonal antibodies. β-Actin was used as internal control. The experiment was repeated three independent times.

Mentions: Bcl-2 family proteins play a key role in the regulation of apoptosis by promoting (Bax and Bak) or inhibiting (Bcl-xL and Bcl-2) cell death. We therefore examined whether Bcl-xL played a role in DPEP-induced apoptosis. Western blot analysis revealed that DPEP treatment suppresses the expression of the anti-apoptotic Bcl-xL protein but did not affect the expression of the pro-apoptotic Bax protein. Further experiments revealed that DPEP induced cleavage of caspase-3 and PARP (Figure 4A(Fig. 4)). The results in Figure 3(Fig. 3) and 4A(Fig. 4) clearly show that both Bcl-xL and ROS are essential for DPEP-induced apoptosis, although the causal relationship between ROS and Bcl-xL is unclear. To further evaluate the significance of ROS generation in DPEP-mediated downregulation of Bcl-xL, HL-60 cells were pretreated with NAC for 1 h, followed by exposure to DPEP for 8 h. As shown in Figure 4B(Fig. 4), pretreatment with NAC attenuated caspase-3 and PARP cleavage and the reduction of Bcl-xL levels.


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)

(A) Effect of DPEP on apoptosis-related protein in HL-60 cells. The cells were exposed to 40 μM DPEP for 1-12 h. (B) Effect of NAC on down-regulation of Bcl-xL and caspase-3, and PARP cleavage activation by DPEP. The cells were pretreated with 1 mM NAC and then treated for 8 h with 40 μM DPEP. Whole cell lysates were subjected to Western blot analysis of anti-Bax, -Bcl-xL, -cleaved-caspase -3, and -PARP monoclonal antibodies. β-Actin was used as internal control. The experiment was repeated three independent times.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834817&req=5

Figure 4: (A) Effect of DPEP on apoptosis-related protein in HL-60 cells. The cells were exposed to 40 μM DPEP for 1-12 h. (B) Effect of NAC on down-regulation of Bcl-xL and caspase-3, and PARP cleavage activation by DPEP. The cells were pretreated with 1 mM NAC and then treated for 8 h with 40 μM DPEP. Whole cell lysates were subjected to Western blot analysis of anti-Bax, -Bcl-xL, -cleaved-caspase -3, and -PARP monoclonal antibodies. β-Actin was used as internal control. The experiment was repeated three independent times.
Mentions: Bcl-2 family proteins play a key role in the regulation of apoptosis by promoting (Bax and Bak) or inhibiting (Bcl-xL and Bcl-2) cell death. We therefore examined whether Bcl-xL played a role in DPEP-induced apoptosis. Western blot analysis revealed that DPEP treatment suppresses the expression of the anti-apoptotic Bcl-xL protein but did not affect the expression of the pro-apoptotic Bax protein. Further experiments revealed that DPEP induced cleavage of caspase-3 and PARP (Figure 4A(Fig. 4)). The results in Figure 3(Fig. 3) and 4A(Fig. 4) clearly show that both Bcl-xL and ROS are essential for DPEP-induced apoptosis, although the causal relationship between ROS and Bcl-xL is unclear. To further evaluate the significance of ROS generation in DPEP-mediated downregulation of Bcl-xL, HL-60 cells were pretreated with NAC for 1 h, followed by exposure to DPEP for 8 h. As shown in Figure 4B(Fig. 4), pretreatment with NAC attenuated caspase-3 and PARP cleavage and the reduction of Bcl-xL levels.

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