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ASK1/JNK-mediated TAp63 activation controls the cell survival signal of baicalein-treated EBV-transformed B cells

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Abstract

Transcriptionally active p63 (TAp63) promotes cell cycle arrest, senescence, and apoptosis in several cancer cells. Migration inhibitory factor (MIF)/CD74 regulates B-cell survival through nuclear factor (NF)-κB-dependent TAp63 expression. In this study, we investigated how the level of TAp63 expression influences the induction of apoptosis in baicalein-treated EBV-transformed B cells. Baicalein induced the expression of TAp63 and apoptosis signal-regulating kinase 1 (ASK1), as well as cytotoxicity, by disrupting the mitochondrial membrane and inhibiting the activation of phosphoinositide 3-kinase (PI3K)/Akt and NF-κB. Genetic knockdown of TAp63 or ASK1 by small interfering RNA resulted in protection from apoptosis accompanied by the recovery of CD74, CD44, α4 integrin, Bcl-2, and NF-κB activation. Baicalein-induced reactive oxygen species activated the ASK1/JNK pathway with subsequent expression of TAp63. Pre-engagement with MIF/CD74 maintained the expression of CD74, CD44, and α4 integrin, as well as Syk/Src-mediated PI3K/Akt activation, in baicalein-treated EBV-transformed B cells. Meanwhile, ASK1/JNK-dependent TAp63 expression was efficiently suppressed after pre-treatment with MIF. Our results suggest that baicalein-mediated ASK1/JNK activation regulates the mitochondria-dependent apoptosis pathway through the up-regulation of TAp63 and down-regulation of NF-κB and CD74/CD44 in B-cell malignancies.

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Abbreviations

7-AAD:

7-Aminoactinomycin D

APDC:

4-Amino-2,4-pyrrolidine-dicarboxylic acid

ASK1:

Apoptosis signal-regulating kinase 1

Cyt. c:

Cytochrome c

DCF:

2979-Dichlorofluorescein

DCFH-DA:

29,79-Dichlorodihydrofluorescein diacetate

DiOC6:

3,39-Dihexyloxacarboxyanine iodide

EBV:

Epstein–Barr virus

EtOH:

Ethanol

GADD34:

Growth arrest and DNA damage-inducible protein 34

GADD45:

Growth arrest and DNA damage protein 45

IP:

Immunoprecipitation

ΔΨm:

Mitochondrial membrane potential

MIF:

Macrophage migration inhibitory factor

NAC:

N-acetylcysteine

PARP:

Poly(ADP-ribose) polymerase

ROS:

Reactive oxygen species

siRNA:

Small interfering RNA

TAp63:

Transcriptionally active p63

TAp73:

Transcriptionally active p73

z-DEVD-fmk:

N-benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethylketone

z-IETDfmk:

N-benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethylketone

z-LEHD-fmk:

z-Leu-Glu(OMe)-His-Asp-(OMe)-fluoromethylketone

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Acknowledgments

This work was supported by the National R&D Program for Cancer Control, Ministry for Health, Welfare and Family Affairs, Republic of Korea (Grant #: 0920040) and Grant from the Korea Healthcare Technology R&D Project of the Ministry of Health and Welfare Affairs, Republic of Korea (Grant#: HI12C0005).

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Authors and Affiliations

  1. Department of Anatomy, Inje University College of Medicine, Busan, 614-735, Republic of Korea

    Ga Bin Park, Yeong Seok Kim, Daejin Kim & Dae Young Hur

  2. Department of Internal Medicine, Inje University Busan Paik Hospital, Busan, 614-735, Republic of Korea

    Hyun-Kyung Lee

  3. Department of Ophthalmology, Inje University Busan Paik Hospital, Busan, Republic of Korea

    Jae Wook Yang

  4. Ocular Neovascular Disease Research Center, Inje University Busan Paik Hospital, Busan, Republic of Korea

    Ga Bin Park, Jae Wook Yang, Daejin Kim & Dae Young Hur

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  1. Ga Bin Park
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Park, G.B., Kim, Y.S., Lee, HK. et al. ASK1/JNK-mediated TAp63 activation controls the cell survival signal of baicalein-treated EBV-transformed B cells. Mol Cell Biochem 412, 247–258 (2016). https://doi.org/10.1007/s11010-015-2631-8

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