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The Study on the Role of JNK/AP-1 and IKK/NF-κB Signaling Pathways in Mediating Arsenite-induced Apoptosis in MCF7 Cells

Author: HaoYi
Tutor: LiWeiPing；SongLun
School: Lanzhou University
Course: Pathology and Pathophysiology
Keywords: As2O3 Breast cancer cells Apoptosis JNK/AP-1 IKK / NF-κB
CLC: R737.9
Type: Master's thesis
Year: 2011
Downloads: 132
Quote: 0
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Abstract


Objective: To detect arsenic trioxide (AS 2 O 3 )-induced apoptosis of breast cancer cells (MCF7) reaction JNK/AP-1 IKK / NF-кB signaling pathway induced activation of to explore mediated AS 2 O 3 -induced breast cancer cell apoptosis reaction of the main signal path. METHODS: Cultured MCF7 cells, AS 2 O 3 source of stimulation, brominated propidium (PI) staining and flow cytometry cell cycle progression and cell apoptotic response; dual luciferase reporter gene assay transcription factor AP-1 and NF-кB induced activation; immunoblot assay JNK, p38K, ERK, MEK1 / 2, c-Jun signaling molecules induced activation and Fra-1, the signaling molecules GADD45a, IKKα, IKKβ, IKKγ of p50, p65 protein expression levels. Results: 1. JNK/AP-1 signaling pathways induced by AS 2 O 3 MCF7 cells apoptosis reaction mechanism of action (1) of the AS 2 O 3 stimulate MCF7 cells G2 / M cell cycle progression arrest and cell apoptosis (2) AS 2 O 3 stimulation show persistent high intensity induced activation of JNK in MCF7 cells. (3) AP-1 transcription activation activity under AS 2 O 3 stimulating effect was dose-and time-dependent upregulation trend, and JNK activation status. (4) AS the 2 O 3 stimulate the next MCF7 cells was significantly activated c-Jun, Fra-1 expression level was significantly upregulated. (5) AP-1 transcriptional activity was inhibited after AS 2 O 3 MCF7 cells induced apoptosis rate significantly reduce. (6) JNK activity was inhibited AS 2 O 3 stimulation transcriptional activation of AP-1 activity in MCF7 cells significantly down-regulated apoptotic response is also significantly reduced. 2. IKK / NF-кB signaling pathway in AS 2 O 3 induced MCF7 cells apoptosis in the role of (1) the AS 2 O < sub> 3 stimulate the transcriptional activation of NF-кB next MCF7 cells showed stimulus dose and time dependent downward trend. (2) AS the 2 O 3 stimulation in MCF7 cells the IKKa and of IKK (protein expression level downward effect of the (3) high expression of IKKa or IKKβ AS 2 O 3 stimulation MCF7 cells, NF-кB transcriptional activity lowering effect was significantly reversed. (4) high expression IKKα or IKKβ detection of AS 2 O 3 stimulation MCF7 cells apoptotic effect was significantly reversed. conclusions: 1. JNK/AP-1 signaling pathway is mediated AS 2 O important signaling pathways induced by 3 apoptotic response of breast cancer cells, AS 2 O 3 has the potential treatment of breast cancer. 2 c-Jun Fra-1 is the MCF7 cells AS 2 O 3 induced activation of the major component of the AP-1 subunit of the AP-1 is located downstream of the JNK pathway mediated AS 2 O 3 key signaling protein molecules that promote apoptotic effects of MCF7 cells, c-Jun, Fra-1 and AP-1 is expected to become a potential intervention target for breast cancer therapy. 3. As 2 O 3 down IKKa and IKKβ expression inhibit the constitutive activation of NF-кB signaling pathways in MCF7 cells, which play survival inhibition and apoptosis role.

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