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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 combined with eIF2α siRNA on cell viability, Caspase 9 activity, ROS induction, NADPH oxidase activity, and p-eIF2α levels in human ESCC cells.(A) Viability is shown as OD values. (B) The intracellular Caspase 9 activity levels are shown. (C) ROS concentrations are shown. (D) NADPH oxidase activity is shown. The two indexes in the control group were defined as 100%. (E) Representative Western blot results of p-eIF2α, CHOP, Bcl2, and PUMA are shown. Membranes were re-probed for β-actin expression to show that similar amounts of protein were loaded in each lane. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control siRNA group; bP < 0.05 vs. the eIF2α siRNA-treated group; cP < 0.05,vs. the control siRNA + ICA 40 μM-treated group.
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f7: Effect of ICA combined with eIF2α siRNA on cell viability, Caspase 9 activity, ROS induction, NADPH oxidase activity, and p-eIF2α levels in human ESCC cells.(A) Viability is shown as OD values. (B) The intracellular Caspase 9 activity levels are shown. (C) ROS concentrations are shown. (D) NADPH oxidase activity is shown. The two indexes in the control group were defined as 100%. (E) Representative Western blot results of p-eIF2α, CHOP, Bcl2, and PUMA are shown. Membranes were re-probed for β-actin expression to show that similar amounts of protein were loaded in each lane. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control siRNA group; bP < 0.05 vs. the eIF2α siRNA-treated group; cP < 0.05,vs. the control siRNA + ICA 40 μM-treated group.

Mentions: eIF2α siRNA was used to explore the effect of downregulating ERS signaling on the antitumor activity of ICA in vitro. ESCC cells were first transfected with eIF2α siRNA and then treated with ICA (40 μM) for an additional 24 h. Transfection with eIF2α siRNA significantly decreased p-eIF2α levels in EC109 and TE1 cells (P < 0.05 compared to transfection with the control siRNA, Fig. 7E). The combination of eIF2α siRNA and ICA significantly increased cell viability (Fig. 7A), decreased Caspase 9 activity (Fig. 7B) and reduced the generation of ROS and NADPH (Fig. 7C,D) (P < 0.05 compared with the combination of control siRNA and ICA); however, eIF2α siRNA alone did not affect cell viability, Caspase 9 activity, ROS generation, or NADPH oxidase activity compared with the control siRNA (P > 0.05). In addition, Bcl2 was further upregulated by co-treatment with ICA and eIF2α siRNA, whereas PUMA was further downregulated by co-treatment with eIF2α siRNA and ICA (P < 0.05 compared with the control siRNA and ICA co-treatment, Fig. 7E).


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 combined with eIF2α siRNA on cell viability, Caspase 9 activity, ROS induction, NADPH oxidase activity, and p-eIF2α levels in human ESCC cells.(A) Viability is shown as OD values. (B) The intracellular Caspase 9 activity levels are shown. (C) ROS concentrations are shown. (D) NADPH oxidase activity is shown. The two indexes in the control group were defined as 100%. (E) Representative Western blot results of p-eIF2α, CHOP, Bcl2, and PUMA are shown. Membranes were re-probed for β-actin expression to show that similar amounts of protein were loaded in each lane. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control siRNA group; bP < 0.05 vs. the eIF2α siRNA-treated group; cP < 0.05,vs. the control siRNA + ICA 40 μM-treated group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f7: Effect of ICA combined with eIF2α siRNA on cell viability, Caspase 9 activity, ROS induction, NADPH oxidase activity, and p-eIF2α levels in human ESCC cells.(A) Viability is shown as OD values. (B) The intracellular Caspase 9 activity levels are shown. (C) ROS concentrations are shown. (D) NADPH oxidase activity is shown. The two indexes in the control group were defined as 100%. (E) Representative Western blot results of p-eIF2α, CHOP, Bcl2, and PUMA are shown. Membranes were re-probed for β-actin expression to show that similar amounts of protein were loaded in each lane. The results are expressed as the mean ± SD; n = 6. aP < 0.05 vs. the control siRNA group; bP < 0.05 vs. the eIF2α siRNA-treated group; cP < 0.05,vs. the control siRNA + ICA 40 μM-treated group.
Mentions: eIF2α siRNA was used to explore the effect of downregulating ERS signaling on the antitumor activity of ICA in vitro. ESCC cells were first transfected with eIF2α siRNA and then treated with ICA (40 μM) for an additional 24 h. Transfection with eIF2α siRNA significantly decreased p-eIF2α levels in EC109 and TE1 cells (P < 0.05 compared to transfection with the control siRNA, Fig. 7E). The combination of eIF2α siRNA and ICA significantly increased cell viability (Fig. 7A), decreased Caspase 9 activity (Fig. 7B) and reduced the generation of ROS and NADPH (Fig. 7C,D) (P < 0.05 compared with the combination of control siRNA and ICA); however, eIF2α siRNA alone did not affect cell viability, Caspase 9 activity, ROS generation, or NADPH oxidase activity compared with the control siRNA (P > 0.05). In addition, Bcl2 was further upregulated by co-treatment with ICA and eIF2α siRNA, whereas PUMA was further downregulated by co-treatment with eIF2α siRNA and ICA (P < 0.05 compared with the control siRNA and ICA co-treatment, Fig. 7E).

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