Ibrutinib, a Bruton’s tyrosine kinase inhibitor, exhibits antitumoral activity and induces autophagy in glioblastomaReportar como inadecuado




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Journal of Experimental and Clinical Cancer Research

, 36:96

Translational Oncology

Abstract

BackgroundGlioblastoma GBM is the most common and aggressive primary brain tumor in adults. Ibrutinib, a Bruton’s tyrosine kinase BTK inhibitor, is a novel anticancer drug used for treating several types of cancers. In this study, we aimed to determine the role of ibrutinib on GBM.

MethodsCell proliferation was determined by using cell viability, colony formation, and 5-ethynyl-2′-deoxyuridine EdU assays. Cell cycle and cell apoptosis were analyzed by flow cytometry. Cell migratory ability was evaluated by wound healing assays and trans-well migration assays. ATG7 expression was knocked-down by transfection with Atg7-specific small interfering RNA. Overexpression of active Akt protein was achieved by transfecting the cells with a plasmid expressing constitutively active Akt CA-Akt. Transmission electron microscopy was performed to examine the formation of autophagosomes in cells. Immunofluorescence and western blot analyses were used to analyze protein expression. Tumor xenografts in nude mice and immunohistochemistry were performed to evaluate the effect of ibrutinib on tumor growth in vivo.

ResultsIbrutinib inhibited cellular proliferation and migration, and induced apoptosis and autophagy in LN229 and U87 cells. Overexpression of the active Akt protein decreased ibrutinib-induced autophagy, while inhibiting Akt by LY294002 treatment enhanced ibrutinib-induced autophagy. Specific inhibition of autophagy by 3-methyladenine 3MA or Atg7 targeting with small interfering RNA si-Atg7 enhanced the anti-GBM effect of ibrutinib in vitro and in vivo.

ConclusionsOur results indicate that ibrutinib exerts a profound antitumor effect and induces autophagy through Akt-mTOR signaling pathway in GBM cells. Autophagy inhibition promotes the antitumor activity of ibrutinib in GBM. Our findings provide important insights into the action of an anticancer agent combining with autophagy inhibitor for malignant glioma.

KeywordsAnti-tumor Autophagy Glioblastoma Ibrutinib Abbreviations3MA3-Methyladenine

AMPKAMP-activated protein kinase

BTKBruton’s tyrosine kinase

CA-AktConstitutively active Akt

CCK-8WST-8 cell counting kit-8

CQChloroquine

EdU5-Ethynyl-2′-deoxyuridine

GBMGlioblastoma

GEOGene expression omnibus

IHCImmunohistochemistry

si-Atg7Small interfering RNA targeting Atg7

TCGAThe Cancer Genome Atlas

TEMTransmission electron microscopy

Electronic supplementary materialThe online version of this article doi:10.1186-s13046-017-0549-6 contains supplementary material, which is available to authorized users.

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Autor: Jin Wang - Xiaoyang Liu - Yongzhi Hong - Songtao Wang - Pin Chen - Aihua Gu - Xiaoyuan Guo - Peng Zhao

Fuente: https://link.springer.com/







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