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Ethanol extract of Forsythia suspensa root induces apoptosis of esophageal carcinoma cells via the mitochondrial apoptotic pathway.

Zhao L, Yan X, Shi J, Ren F, Liu L, Sun S, Shan B - Mol Med Rep (2014)

Bottom Line: The results revealed that FSREE, rather than Forsythia suspensa ethanolic extracts from the leaf (FSLEE) and fruit (FSFEE) exhibited marked anti-tumor activity towards human esophageal cancer cells.FSREE induced cancer cell apoptosis and growth arrest by downregulating B‑cell lymphoma (Bcl)‑2, Bcl‑extra large and myeloid cell leukemia 1, while upregulating Bcl‑2‑associated X protein, Bcl‑2 antagonist of cell death and phorbol‑12‑myristate‑13‑acetate‑induced protein 1.Furthermore, the anti-cancer activity of FSREE was associated with a decreased level of phosphorylated Janus kinase/signal transducer and activator of transcription 3 and extracellular‑signal‑regulated kinase signaling activity.

View Article: PubMed Central - PubMed

Affiliation: Research Center, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.

ABSTRACT
Forsythia suspensa root is used in the treatment of fever and jaundice in Traditional Chinese Medicine. In the present study, the anti-tumor activity of the ethanolic extract of Forsythia suspensa root (FSREE) against esophageal carcinoma cells was investigated in vitro and in vivo and its anti-cancer mechanism was examined. The results revealed that FSREE, rather than Forsythia suspensa ethanolic extracts from the leaf (FSLEE) and fruit (FSFEE) exhibited marked anti-tumor activity towards human esophageal cancer cells. FSREE induced cancer cell apoptosis and growth arrest by downregulating B‑cell lymphoma (Bcl)‑2, Bcl‑extra large and myeloid cell leukemia 1, while upregulating Bcl‑2‑associated X protein, Bcl‑2 antagonist of cell death and phorbol‑12‑myristate‑13‑acetate‑induced protein 1. This led to the activation of poly(ADP ribose) polymerase, caspase‑3 and caspase‑9, but not caspase‑8. Furthermore, the anti-cancer activity of FSREE was associated with a decreased level of phosphorylated Janus kinase/signal transducer and activator of transcription 3 and extracellular‑signal‑regulated kinase signaling activity. It was also observed that the levels of cytochrome c were elevated in the cytoplasm, accounting for the loss of mitochondrial membrane potential in the TE‑13 cells upon treatment with FSEER. In addition, FSEER inhibited the growth of esophageal cancer cells in xenograft models and no detectable toxicity was present in the lung or liver tissues. These observations provided further evidence of the anti-tumor effect of FSEER and may be of importance to further examine the potential role of Forsythia suspensa root as a therapeutic agent in esophageal carcinoma therapy.

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Inactivity of FSEER on the JAK/STAT3 and ERK signaling pathways in TE-13 cells. (A) TE-13 cells were treated with FSEER at 0.25, 0.5 and 1.0 mg/ml for 60 min and with (B) 0.5 mg/ml FSEER for 15, 30 and 60 min. (C) Effects of AG490 (5 mol/l) or PD98059 (10 μmol/l) alone or in combination with FSEER on the FSEER-mediated TE-13 cell proliferation. *P<0.05, compared with the control group. FSEER, Forsythia suspensa ethanolic extract of the root; p-JAK, phosphorylated Janus kinase; STAT, signal transducer and activator of transcription; ERK, extracellular-signal-regulated kinase.
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f5-mmr-11-02-0871: Inactivity of FSEER on the JAK/STAT3 and ERK signaling pathways in TE-13 cells. (A) TE-13 cells were treated with FSEER at 0.25, 0.5 and 1.0 mg/ml for 60 min and with (B) 0.5 mg/ml FSEER for 15, 30 and 60 min. (C) Effects of AG490 (5 mol/l) or PD98059 (10 μmol/l) alone or in combination with FSEER on the FSEER-mediated TE-13 cell proliferation. *P<0.05, compared with the control group. FSEER, Forsythia suspensa ethanolic extract of the root; p-JAK, phosphorylated Janus kinase; STAT, signal transducer and activator of transcription; ERK, extracellular-signal-regulated kinase.

Mentions: The JAK/STAT3 and ERK signaling pathways are important pathways in cell growth and apoptosis and the inactivity of these pathways may regulate the Bcl2 family resulting in growth arrest and apoptosis in certain tumor cells (19–21). Several studies have suggested that anti-apoptotic genes are regulated by interleukin 6 and STAT3, including Bcl-2, Bcl-xL and Mcl-1 (22). While these genes are induced by STAT3, the most important anti-apoptotic gene is considered to be Mcl-1 and Bcl-xL (23). The results of the present study revealed that FSEER markedly reduced the expression of p-JAK/STAT3 and p-ERK in a concentration and time-dependent manner (Fig. 5A and B), indicating that FSEER inhibited the activation of the JAK/STAT3 and ERK signaling pathways in TE-13 cells. In order to verify the involvement of these pathways in FSEER-induced apoptosis, the effect of FSEER on the proliferation of TE-13 cells was observed in the presence of an inhibitor of the signaling pathway. AG490 is a member of the typhostin family of tyrosine kinase inhibitors, which inhibit the JAK/STAT3 signaling pathway in several types of cancer cell, including esophageal carcinoma cells (24,25). Beales and Ogunwobi (26) demonstrated that the P4244 MAP kinase inhibitor PD98059 enhanced the activity of leptin-mediated esophageal adenocarcinoma cell apoptosis. The present study revealed that, although AG490 (5 μmol/l) and PD98059 (10 μmol/l) alone were not able to inhibit the proliferation of TE-13 cells, they significantly enhanced the inhibitory effect of FSEER (0.5 mg/ml) on the proliferation of TE-13 cells by ~25 and 35%, respectively (Fig. 5C). Taken together, the findings of the present study demonstrated that the induction of apoptosis of TE-13 cells by FSEER was achieved through downregulation of the JAK/STAT3 and ERK signaling pathways.


Ethanol extract of Forsythia suspensa root induces apoptosis of esophageal carcinoma cells via the mitochondrial apoptotic pathway.

Zhao L, Yan X, Shi J, Ren F, Liu L, Sun S, Shan B - Mol Med Rep (2014)

Inactivity of FSEER on the JAK/STAT3 and ERK signaling pathways in TE-13 cells. (A) TE-13 cells were treated with FSEER at 0.25, 0.5 and 1.0 mg/ml for 60 min and with (B) 0.5 mg/ml FSEER for 15, 30 and 60 min. (C) Effects of AG490 (5 mol/l) or PD98059 (10 μmol/l) alone or in combination with FSEER on the FSEER-mediated TE-13 cell proliferation. *P<0.05, compared with the control group. FSEER, Forsythia suspensa ethanolic extract of the root; p-JAK, phosphorylated Janus kinase; STAT, signal transducer and activator of transcription; ERK, extracellular-signal-regulated kinase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262507&req=5

f5-mmr-11-02-0871: Inactivity of FSEER on the JAK/STAT3 and ERK signaling pathways in TE-13 cells. (A) TE-13 cells were treated with FSEER at 0.25, 0.5 and 1.0 mg/ml for 60 min and with (B) 0.5 mg/ml FSEER for 15, 30 and 60 min. (C) Effects of AG490 (5 mol/l) or PD98059 (10 μmol/l) alone or in combination with FSEER on the FSEER-mediated TE-13 cell proliferation. *P<0.05, compared with the control group. FSEER, Forsythia suspensa ethanolic extract of the root; p-JAK, phosphorylated Janus kinase; STAT, signal transducer and activator of transcription; ERK, extracellular-signal-regulated kinase.
Mentions: The JAK/STAT3 and ERK signaling pathways are important pathways in cell growth and apoptosis and the inactivity of these pathways may regulate the Bcl2 family resulting in growth arrest and apoptosis in certain tumor cells (19–21). Several studies have suggested that anti-apoptotic genes are regulated by interleukin 6 and STAT3, including Bcl-2, Bcl-xL and Mcl-1 (22). While these genes are induced by STAT3, the most important anti-apoptotic gene is considered to be Mcl-1 and Bcl-xL (23). The results of the present study revealed that FSEER markedly reduced the expression of p-JAK/STAT3 and p-ERK in a concentration and time-dependent manner (Fig. 5A and B), indicating that FSEER inhibited the activation of the JAK/STAT3 and ERK signaling pathways in TE-13 cells. In order to verify the involvement of these pathways in FSEER-induced apoptosis, the effect of FSEER on the proliferation of TE-13 cells was observed in the presence of an inhibitor of the signaling pathway. AG490 is a member of the typhostin family of tyrosine kinase inhibitors, which inhibit the JAK/STAT3 signaling pathway in several types of cancer cell, including esophageal carcinoma cells (24,25). Beales and Ogunwobi (26) demonstrated that the P4244 MAP kinase inhibitor PD98059 enhanced the activity of leptin-mediated esophageal adenocarcinoma cell apoptosis. The present study revealed that, although AG490 (5 μmol/l) and PD98059 (10 μmol/l) alone were not able to inhibit the proliferation of TE-13 cells, they significantly enhanced the inhibitory effect of FSEER (0.5 mg/ml) on the proliferation of TE-13 cells by ~25 and 35%, respectively (Fig. 5C). Taken together, the findings of the present study demonstrated that the induction of apoptosis of TE-13 cells by FSEER was achieved through downregulation of the JAK/STAT3 and ERK signaling pathways.

Bottom Line: The results revealed that FSREE, rather than Forsythia suspensa ethanolic extracts from the leaf (FSLEE) and fruit (FSFEE) exhibited marked anti-tumor activity towards human esophageal cancer cells.FSREE induced cancer cell apoptosis and growth arrest by downregulating B‑cell lymphoma (Bcl)‑2, Bcl‑extra large and myeloid cell leukemia 1, while upregulating Bcl‑2‑associated X protein, Bcl‑2 antagonist of cell death and phorbol‑12‑myristate‑13‑acetate‑induced protein 1.Furthermore, the anti-cancer activity of FSREE was associated with a decreased level of phosphorylated Janus kinase/signal transducer and activator of transcription 3 and extracellular‑signal‑regulated kinase signaling activity.

View Article: PubMed Central - PubMed

Affiliation: Research Center, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.

ABSTRACT
Forsythia suspensa root is used in the treatment of fever and jaundice in Traditional Chinese Medicine. In the present study, the anti-tumor activity of the ethanolic extract of Forsythia suspensa root (FSREE) against esophageal carcinoma cells was investigated in vitro and in vivo and its anti-cancer mechanism was examined. The results revealed that FSREE, rather than Forsythia suspensa ethanolic extracts from the leaf (FSLEE) and fruit (FSFEE) exhibited marked anti-tumor activity towards human esophageal cancer cells. FSREE induced cancer cell apoptosis and growth arrest by downregulating B‑cell lymphoma (Bcl)‑2, Bcl‑extra large and myeloid cell leukemia 1, while upregulating Bcl‑2‑associated X protein, Bcl‑2 antagonist of cell death and phorbol‑12‑myristate‑13‑acetate‑induced protein 1. This led to the activation of poly(ADP ribose) polymerase, caspase‑3 and caspase‑9, but not caspase‑8. Furthermore, the anti-cancer activity of FSREE was associated with a decreased level of phosphorylated Janus kinase/signal transducer and activator of transcription 3 and extracellular‑signal‑regulated kinase signaling activity. It was also observed that the levels of cytochrome c were elevated in the cytoplasm, accounting for the loss of mitochondrial membrane potential in the TE‑13 cells upon treatment with FSEER. In addition, FSEER inhibited the growth of esophageal cancer cells in xenograft models and no detectable toxicity was present in the lung or liver tissues. These observations provided further evidence of the anti-tumor effect of FSEER and may be of importance to further examine the potential role of Forsythia suspensa root as a therapeutic agent in esophageal carcinoma therapy.

Show MeSH
Related in: MedlinePlus