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Icariin displays anticancer activity against human esophageal cancer cells via regulating endoplasmic reticulum stress-mediated apoptotic signaling.

Fan C, Yang Y, Liu Y, Jiang S, Di S, Hu W, Ma Z, Li T, Zhu Y, Xin Z, Wu G, Han J, Li X, Yan X - Sci Rep (2016)

Bottom Line: ICA treatment resulted in a dose- and time-dependent decrease in the viability of human EC109 and TE1 ESCCs.The downregulation of ERS signaling using eIF2α siRNA desensitized EC109 and TE1 cells to ICA treatment, and the upregulation of ERS signaling using thapsigargin sensitized EC109 and TE1 cells to ICA treatment.In summary, ERS activation may represent a mechanism of action for the anticancer activity of ICA in ESCCs, and the activation of ERS signaling may represent a novel therapeutic intervention for human esophageal cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, 1 Xinsi Road, Xi'an 710038, China.

ABSTRACT
In this study, we investigated the antitumor activity of icariin (ICA) in human esophageal squamous cell carcinoma (ESCC) in vitro and in vivo and explored the role of endoplasmic reticulum stress (ERS) signaling in this activity. ICA treatment resulted in a dose- and time-dependent decrease in the viability of human EC109 and TE1 ESCCs. Additionally, ICA exhibited strong antitumor activity, as evidenced by reductions in cell migration, adhesion, and intracellular glutathione (GSH) levels and by increases in the EC109 and TE1 cell apoptotic index, Caspase 9 activity, reactive oxygen species (ROS) level, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity. Furthermore, ICA treatments upregulated the levels of ERS-related molecules (p-PERK, GRP78, ATF4, p-eIF2α, and CHOP) and a pro-apoptotic protein (PUMA) and simultaneously downregulated an anti-apoptotic protein (Bcl2) in the two ESCC cell lines. The downregulation of ERS signaling using eIF2α siRNA desensitized EC109 and TE1 cells to ICA treatment, and the upregulation of ERS signaling using thapsigargin sensitized EC109 and TE1 cells to ICA treatment. In summary, ERS activation may represent a mechanism of action for the anticancer activity of ICA in ESCCs, and the activation of ERS signaling may represent a novel therapeutic intervention for human esophageal cancer.

No MeSH data available.


Related in: MedlinePlus

Effect of ICA treatment on Caspase 9 activity, ROS generation, NADPH oxidase activity, and GSH levels in human ESCC cells (24 h).(A) The intracellular Caspase 9 activity levels are shown. (B) ROS concentrations are shown. (C) NADPH oxidase activity is shown. (D) Intracellular GSH levels are shown. The three indexes in the control group were defined as 100%. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control group; bP < 0.05 vs. the 20 μM ICA-treated group; cP < 0.05 vs. the 40 μM ICA-treated group.
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f5: Effect of ICA treatment on Caspase 9 activity, ROS generation, NADPH oxidase activity, and GSH levels in human ESCC cells (24 h).(A) The intracellular Caspase 9 activity levels are shown. (B) ROS concentrations are shown. (C) NADPH oxidase activity is shown. (D) Intracellular GSH levels are shown. The three indexes in the control group were defined as 100%. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control group; bP < 0.05 vs. the 20 μM ICA-treated group; cP < 0.05 vs. the 40 μM ICA-treated group.

Mentions: After ICA treatment (20, 40, or 80 μM), Caspase 9 activity (Fig. 5A) was significantly increased by 346.84 ± 24.58%, 426.52 ± 27.21%, and 489.23 ± 26.39% in EC109 cells and 321.98 ± 22.56%, 419.21 ± 25.67%, and 492.63 ± 27.66% in TE1 cells, respectively (P < 0.05 compared with the control treatment). Next, intracellular ROS production was analyzed based on the ROS-mediated conversion of the non-fluorescent species 2’,7’-DCFH-DA to the fluorescent species DCFH, which exhibits enhanced fluorescence intensity following the generation of intracellular reactive metabolites. Treatment with ICA (20, 40, or 80 μM) for 24 h induced a dose-dependent increase in ROS generation by 216.46 ± 23.57%, 336.80 ± 25.11%, and 451.80 ± 23.63% in EC109 cells and 224.46 ± 24.67%, 349.80 ± 23.15%, and 457.81 ± 25.18% in TE1 cells, respectively (P < 0.05 compared with the control treatment) (Fig. 5B). The NADPH oxidase system is now widely recognized as a key player in intracellular ROS homeostasis and as one of the major producers of ROS within the cell32. After administrated with ICA as above doses, NADPH oxidase activity was increased at a dose-dependent manner from 183.46 ± 25.68%, 269.37 ± 22.58% to 307.67 ± 22.15% in EC109 cells and 192.52 ± 22.86%, 25.82 ± 22.28% to 338.17 ± 3.12% in TE1 cells, respectively (P < 0.05 compared with the control treatment) (Fig. 5C). GSH is the major non-protein thiol in cells and is essential for maintaining cellular redox homeostasis. After treatment with ICA (20, 40, or 80 μM) for 24 h, a dose-dependent decrease in the intracellular GSH levels (to 71.29 ± 6.37%, 58.74 ± 6.98%, and 36.78 ± 8.91% of the levels in controls in EC109 cells and 68.29 ± 5.51%, 56.97 ± 6.84%, and 34.28 ± 8.93% of the levels in controls in TE1 cells, respectively) was observed (P < 0.05 compared with the control treatment, Fig. 5D).


Icariin displays anticancer activity against human esophageal cancer cells via regulating endoplasmic reticulum stress-mediated apoptotic signaling.

Fan C, Yang Y, Liu Y, Jiang S, Di S, Hu W, Ma Z, Li T, Zhu Y, Xin Z, Wu G, Han J, Li X, Yan X - Sci Rep (2016)

Effect of ICA treatment on Caspase 9 activity, ROS generation, NADPH oxidase activity, and GSH levels in human ESCC cells (24 h).(A) The intracellular Caspase 9 activity levels are shown. (B) ROS concentrations are shown. (C) NADPH oxidase activity is shown. (D) Intracellular GSH levels are shown. The three indexes in the control group were defined as 100%. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control group; bP < 0.05 vs. the 20 μM ICA-treated group; cP < 0.05 vs. the 40 μM ICA-treated group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4759694&req=5

f5: Effect of ICA treatment on Caspase 9 activity, ROS generation, NADPH oxidase activity, and GSH levels in human ESCC cells (24 h).(A) The intracellular Caspase 9 activity levels are shown. (B) ROS concentrations are shown. (C) NADPH oxidase activity is shown. (D) Intracellular GSH levels are shown. The three indexes in the control group were defined as 100%. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control group; bP < 0.05 vs. the 20 μM ICA-treated group; cP < 0.05 vs. the 40 μM ICA-treated group.
Mentions: After ICA treatment (20, 40, or 80 μM), Caspase 9 activity (Fig. 5A) was significantly increased by 346.84 ± 24.58%, 426.52 ± 27.21%, and 489.23 ± 26.39% in EC109 cells and 321.98 ± 22.56%, 419.21 ± 25.67%, and 492.63 ± 27.66% in TE1 cells, respectively (P < 0.05 compared with the control treatment). Next, intracellular ROS production was analyzed based on the ROS-mediated conversion of the non-fluorescent species 2’,7’-DCFH-DA to the fluorescent species DCFH, which exhibits enhanced fluorescence intensity following the generation of intracellular reactive metabolites. Treatment with ICA (20, 40, or 80 μM) for 24 h induced a dose-dependent increase in ROS generation by 216.46 ± 23.57%, 336.80 ± 25.11%, and 451.80 ± 23.63% in EC109 cells and 224.46 ± 24.67%, 349.80 ± 23.15%, and 457.81 ± 25.18% in TE1 cells, respectively (P < 0.05 compared with the control treatment) (Fig. 5B). The NADPH oxidase system is now widely recognized as a key player in intracellular ROS homeostasis and as one of the major producers of ROS within the cell32. After administrated with ICA as above doses, NADPH oxidase activity was increased at a dose-dependent manner from 183.46 ± 25.68%, 269.37 ± 22.58% to 307.67 ± 22.15% in EC109 cells and 192.52 ± 22.86%, 25.82 ± 22.28% to 338.17 ± 3.12% in TE1 cells, respectively (P < 0.05 compared with the control treatment) (Fig. 5C). GSH is the major non-protein thiol in cells and is essential for maintaining cellular redox homeostasis. After treatment with ICA (20, 40, or 80 μM) for 24 h, a dose-dependent decrease in the intracellular GSH levels (to 71.29 ± 6.37%, 58.74 ± 6.98%, and 36.78 ± 8.91% of the levels in controls in EC109 cells and 68.29 ± 5.51%, 56.97 ± 6.84%, and 34.28 ± 8.93% of the levels in controls in TE1 cells, respectively) was observed (P < 0.05 compared with the control treatment, Fig. 5D).

Bottom Line: ICA treatment resulted in a dose- and time-dependent decrease in the viability of human EC109 and TE1 ESCCs.The downregulation of ERS signaling using eIF2α siRNA desensitized EC109 and TE1 cells to ICA treatment, and the upregulation of ERS signaling using thapsigargin sensitized EC109 and TE1 cells to ICA treatment.In summary, ERS activation may represent a mechanism of action for the anticancer activity of ICA in ESCCs, and the activation of ERS signaling may represent a novel therapeutic intervention for human esophageal cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, 1 Xinsi Road, Xi'an 710038, China.

ABSTRACT
In this study, we investigated the antitumor activity of icariin (ICA) in human esophageal squamous cell carcinoma (ESCC) in vitro and in vivo and explored the role of endoplasmic reticulum stress (ERS) signaling in this activity. ICA treatment resulted in a dose- and time-dependent decrease in the viability of human EC109 and TE1 ESCCs. Additionally, ICA exhibited strong antitumor activity, as evidenced by reductions in cell migration, adhesion, and intracellular glutathione (GSH) levels and by increases in the EC109 and TE1 cell apoptotic index, Caspase 9 activity, reactive oxygen species (ROS) level, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity. Furthermore, ICA treatments upregulated the levels of ERS-related molecules (p-PERK, GRP78, ATF4, p-eIF2α, and CHOP) and a pro-apoptotic protein (PUMA) and simultaneously downregulated an anti-apoptotic protein (Bcl2) in the two ESCC cell lines. The downregulation of ERS signaling using eIF2α siRNA desensitized EC109 and TE1 cells to ICA treatment, and the upregulation of ERS signaling using thapsigargin sensitized EC109 and TE1 cells to ICA treatment. In summary, ERS activation may represent a mechanism of action for the anticancer activity of ICA in ESCCs, and the activation of ERS signaling may represent a novel therapeutic intervention for human esophageal cancer.

No MeSH data available.


Related in: MedlinePlus