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New benzimidazole acridine derivative induces human colon cancer cell apoptosis in vitro via the ROS-JNK signaling pathway.

Chen K, Chu BZ, Liu F, Li B, Gao CM, Li LL, Sun QS, Shen ZF, Jiang YY - Acta Pharmacol. Sin. (2015)

Bottom Line: Knockout of JNK1, but not p38, attenuated 8m-induced apoptosis.In addition, 8m induced ROS production and mitochondrial membrane depolarization in HCT116 cells.Pretreatment with the antioxidants N-acetyl cysteine or glutathione attenuated 8m-induced apoptosis and JNK activation in HCT116 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Tsinghua University, Beijing 100084, China.

ABSTRACT

Aim: To investigate the mechanisms underlying anticancer action of the benzimidazole acridine derivative N-{(1H-benzo[d]imidazol-2-yl)methyl}-2-butylacridin-9-amine(8m) against human colon cancer cells in vitro.

Methods: Human colon cancer cell lines SW480 and HCT116 were incubated in the presence of 8m, and then the cell proliferation and apoptosis were measured. The expression of apoptotic/signaling genes and proteins was detected using RT-PCR and Western blotting. ROS generation and mitochondrial membrane depolarization were visualized with fluorescence microscopy.

Results: 8m dose-dependently suppressed the proliferation of SW480 and HCT116 cells with IC50 values of 6.77 and 3.33 μmol/L, respectively. 8m induced apoptosis of HCT116 cells, accompanied by down-regulation of Bcl-2, up-regulation of death receptor-5 (DR5), truncation of Bid, cleavage of PARP, and activation of caspases (including caspase-8 and caspase-9 as well as the downstream caspases-3 and caspase-7). Moreover, 8m selectively activated JNK and p38 without affecting ERK in HCT116 cells. Knockout of JNK1, but not p38, attenuated 8m-induced apoptosis. In addition, 8m induced ROS production and mitochondrial membrane depolarization in HCT116 cells. Pretreatment with the antioxidants N-acetyl cysteine or glutathione attenuated 8m-induced apoptosis and JNK activation in HCT116 cells.

Conclusion: The new benzimidazole acridine derivative, 8m exerts anticancer activity against human colon cancer cells in vitro by inducing both intrinsic and extrinsic apoptosis pathways via the ROS-JNK1 pathway.

No MeSH data available.


Related in: MedlinePlus

Effects of 8m on MAPK activation. Phosphorylation of JNK, p38 MAP kinase, and ERK in cell lysates was detected by immunoblot analysis using the indicated antibodies. (A) HCT116 cells were treated with the indicated concentrations of 8m for 24 h. (B) HCT116 cells were treated with 5 μmol/L 8m for 4, 8, 12, or 24 h.
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fig3: Effects of 8m on MAPK activation. Phosphorylation of JNK, p38 MAP kinase, and ERK in cell lysates was detected by immunoblot analysis using the indicated antibodies. (A) HCT116 cells were treated with the indicated concentrations of 8m for 24 h. (B) HCT116 cells were treated with 5 μmol/L 8m for 4, 8, 12, or 24 h.

Mentions: MAPK signaling can be activated in response to a variety of extracellular stimuli, including mitogens, heat shock and osmotic stress, and can regulate the levels of DRs, Bcl-2, and other apoptosis-related proteins36,37,38. To determine whether MAPK is involved in apoptosis after 8m treatment, we examined the effects of 8m on the activation of MAPK pathways. Treatment of HCT116 cells with 8m increased the amounts of phosphorylated JNK and p38, which reflect the activated states of these two kinases, in a dose-dependent manner. However, the phosphorylation of ERK was minimally affected (Figure 3A). Our subsequent analysis of the time course of JNK and p38 activation showed that the phosphorylation of JNK and p38 were time-dependent: both were induced at 4 h after exposure to 8m, reaching a maximum level at 12 h, with activity persisting until 24 h. Fluctuations in levels of phosphorylated ERK were barely apparent during this time period (Figure 3B). In addition, we noted that c-Fos, a well-known JNK substrate, was also up-regulated by 8m treatment in the HCT116 colon cancer cell line (P<0.05, Figure 2A).


New benzimidazole acridine derivative induces human colon cancer cell apoptosis in vitro via the ROS-JNK signaling pathway.

Chen K, Chu BZ, Liu F, Li B, Gao CM, Li LL, Sun QS, Shen ZF, Jiang YY - Acta Pharmacol. Sin. (2015)

Effects of 8m on MAPK activation. Phosphorylation of JNK, p38 MAP kinase, and ERK in cell lysates was detected by immunoblot analysis using the indicated antibodies. (A) HCT116 cells were treated with the indicated concentrations of 8m for 24 h. (B) HCT116 cells were treated with 5 μmol/L 8m for 4, 8, 12, or 24 h.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4561968&req=5

fig3: Effects of 8m on MAPK activation. Phosphorylation of JNK, p38 MAP kinase, and ERK in cell lysates was detected by immunoblot analysis using the indicated antibodies. (A) HCT116 cells were treated with the indicated concentrations of 8m for 24 h. (B) HCT116 cells were treated with 5 μmol/L 8m for 4, 8, 12, or 24 h.
Mentions: MAPK signaling can be activated in response to a variety of extracellular stimuli, including mitogens, heat shock and osmotic stress, and can regulate the levels of DRs, Bcl-2, and other apoptosis-related proteins36,37,38. To determine whether MAPK is involved in apoptosis after 8m treatment, we examined the effects of 8m on the activation of MAPK pathways. Treatment of HCT116 cells with 8m increased the amounts of phosphorylated JNK and p38, which reflect the activated states of these two kinases, in a dose-dependent manner. However, the phosphorylation of ERK was minimally affected (Figure 3A). Our subsequent analysis of the time course of JNK and p38 activation showed that the phosphorylation of JNK and p38 were time-dependent: both were induced at 4 h after exposure to 8m, reaching a maximum level at 12 h, with activity persisting until 24 h. Fluctuations in levels of phosphorylated ERK were barely apparent during this time period (Figure 3B). In addition, we noted that c-Fos, a well-known JNK substrate, was also up-regulated by 8m treatment in the HCT116 colon cancer cell line (P<0.05, Figure 2A).

Bottom Line: Knockout of JNK1, but not p38, attenuated 8m-induced apoptosis.In addition, 8m induced ROS production and mitochondrial membrane depolarization in HCT116 cells.Pretreatment with the antioxidants N-acetyl cysteine or glutathione attenuated 8m-induced apoptosis and JNK activation in HCT116 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Tsinghua University, Beijing 100084, China.

ABSTRACT

Aim: To investigate the mechanisms underlying anticancer action of the benzimidazole acridine derivative N-{(1H-benzo[d]imidazol-2-yl)methyl}-2-butylacridin-9-amine(8m) against human colon cancer cells in vitro.

Methods: Human colon cancer cell lines SW480 and HCT116 were incubated in the presence of 8m, and then the cell proliferation and apoptosis were measured. The expression of apoptotic/signaling genes and proteins was detected using RT-PCR and Western blotting. ROS generation and mitochondrial membrane depolarization were visualized with fluorescence microscopy.

Results: 8m dose-dependently suppressed the proliferation of SW480 and HCT116 cells with IC50 values of 6.77 and 3.33 μmol/L, respectively. 8m induced apoptosis of HCT116 cells, accompanied by down-regulation of Bcl-2, up-regulation of death receptor-5 (DR5), truncation of Bid, cleavage of PARP, and activation of caspases (including caspase-8 and caspase-9 as well as the downstream caspases-3 and caspase-7). Moreover, 8m selectively activated JNK and p38 without affecting ERK in HCT116 cells. Knockout of JNK1, but not p38, attenuated 8m-induced apoptosis. In addition, 8m induced ROS production and mitochondrial membrane depolarization in HCT116 cells. Pretreatment with the antioxidants N-acetyl cysteine or glutathione attenuated 8m-induced apoptosis and JNK activation in HCT116 cells.

Conclusion: The new benzimidazole acridine derivative, 8m exerts anticancer activity against human colon cancer cells in vitro by inducing both intrinsic and extrinsic apoptosis pathways via the ROS-JNK1 pathway.

No MeSH data available.


Related in: MedlinePlus