Dasatinib promotes paclitaxel-induced necroptosis in lung adenocarcinoma with phosphorylated caspase-8 by c-Src

doi:10.1016/j.canlet.2016.05.003

Cancer Letters

Volume 379, Issue 1, 28 August 2016, Pages 12-23

https://doi.org/10.1016/j.canlet.2016.05.003 Get rights and content

Highlights

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Phosphorylated caspase-8 by c-Src indicated lower response rate to TP regimen in patients with lung adenocarcinoma.
•
Paclitaxel induced necroptosis in lung adenocarcinoma cells dependently of c-Src-phosphorylated caspase-8, RIPK1 and RIPK3.
•
c-Src inhibitor dephosphorylated caspase-8 to promote necroptosis in the paclitaxel-treated Casp8⁺Src⁺ cells.

Abstract

Cisplatin and paclitaxel are considered to be the backbone of chemotherapy in lung adenocarcinoma. These agents show pleiotropic effects on cell death. However, the precise mechanisms remain unclear. The present study reported that phosphorylated caspase-8 at tyrosine 380 (p-Casp8) was characterized as a biomarker of chemoresistance to TP regimen (cisplatin and paclitaxel) in patients with resectable lung adenocarcinoma with significantly poorer 5-year disease-free survival (DFS) and overall survival (OS). Cisplatin killed lung adenocarcinoma cells regardless of c-Src-induced caspase-8 phosphorylation at tyrosine 380. Subsequently, we identified a novel mechanism by which paclitaxel induced necroptosis in lung adenocarcinoma cells that was dependent upon p-Casp8, receptor-interacting protein kinase 1 (RIPK1), and RIPK3. Moreover, dasatinib, a c-Src inhibitor, dephosphorylated caspase-8 to facilitate necroptosis, rather than apoptosis, in paclitaxel-treated p-Casp8-expressing lung adenocarcinoma cells. The data from our study revealed previously unrecognized roles of p-Casp8 as a positive effector in the initiation of necroptosis and as a negative effector in the repression of the interaction between RIPK1 and RIPK3. Moreover, these outcomes supported the need for further clinical studies with the goal of evaluating the efficacy of dasatinib plus paclitaxel in the treatment of lung adenocarcinoma.

Introduction

During the last decade, lung cancer has become the leading cause of cancer-related deaths with a 5-year overall survival (OS) of 15% worldwide [1]. Non-small cell lung cancer (NSCLC) accounts for 80%–85% of all lung cancers, with lung adenocarcinoma representing the largest fraction of NSCLC. A large number of patients with resectable lung adenocarcinoma who respond initially to chemotherapy will subsequently progress [2], [3]. Therefore, identifying molecular determinants of resistance to chemotherapy in lung adenocarcinomas is of paramount importance to improve the clinical efficacy of current treatment regimens.

Cisplatin and paclitaxel are currently considered to be the backbone of chemotherapy for lung adenocarcinoma [2], [3], [4]. Although the primary pharmacological mechanisms of antitumor drugs vary, it has become evident that antitumor drug-induced cell killing is characterized by programmed cell death (apoptosis) that is highly conserved across various species [5]. Aside from apoptosis, another form of programmed cell death, necroptosis, has been discovered more recently [5], [6]. Necroptosis has been shown to occur only upon the pharmacological inhibition or genetic ablation of apoptotic pathways, suggesting that necroptosis is induced as an alternative to ensure cell death when apoptosis fails [5], [6], [7]. Furthermore, c-Src, a non-receptor tyrosine kinase (NRTK), was demonstrated to phosphorylate caspase-8 on tyrosine 380 (p-Casp8). This specific phosphorylation impairs caspase-8 cleavage in the apoptotic cascade [8]. Whether caspase-8 functions as a decision point that directs cell death to apoptosis or necroptosis remains unclear. In our study, cisplatin killed lung adenocarcinoma cells in a manner that was independent of p-Casp8. We uncovered a death receptor-independent necroptotic pathway following paclitaxel treatment that resulted in the phosphorylation of caspase-8 by c-Src and the initiation of necroptosis through recruitment and activation of RIPK1 and RIPK3. It was intriguing that dasatinib, a c-Src inhibitor, markedly facilitated paclitaxel-triggered necroptosis in p-Casp8-expressing lung adenocarcinoma cells via caspase-8 dephosphorylation.

Section snippets

Patients and chemotherapy

Human lung adenocarcinoma and adjacent paracancerous tissues (≥2.0 cm from primary tumor site) from 109 patients were collected following surgeries at the Department of Pathology of the First and Second Affiliated Hospital of Xi'an Jiaotong University from 2006 to 2009. Lung adenocarcinoma was determined by two individual pathologists and classified as pathological stage I-IIIA according to AJCC 2013. The eligible patients received two cycles of neoadjuvant chemotherapy: 135 mg of paclitaxel

p-Casp8 was predictive of a poorer prognosis in patients with resectable lung adenocarcinoma

To screen for prognostic indicators in patients with resectable lung adenocarcinoma, protein extracts from 109 cancerous and paracancerous tissues were incubated with protein microarrays containing monoclonal antibodies against 500 human proteins. Our prior study had confirmed that caspase-8/p-Casp8 and c-Src/p-Src were expressed in A549 cells [8]. Herein, the protein extract of A549 cells was used as a positive control to assess the relative expression levels of

Discussion

Numerous studies have identified differences in gene expression between cancerous and paracancerous tissues using such techniques as serial analysis of gene expression and cDNA microarrays [23], [24]. However, the levels of mRNAs do not always reflect the accurate levels of corresponding proteins, nor do they reveal changes in the post-transcriptional modification of proteins or changes in protein degradation rates. Therefore, it is important to analyze differences in protein levels as a

Funding

Supported by National Natural Science Foundation of China (No. 81301847) and the Fundamental Research Funds (xjj2013071) for the Central Universities, and Shaanxi Province Science and Technology Research Development Program (2014K-01-01-22).

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Original ArticlesDasatinib promotes paclitaxel-induced necroptosis in lung adenocarcinoma with phosphorylated caspase-8 by c-Src

Highlights

Abstract

Introduction

Section snippets

Patients and chemotherapy

p-Casp8 was predictive of a poorer prognosis in patients with resectable lung adenocarcinoma

Discussion

Funding

Biochem. Pharmacol

Cell

Cell

Cell. Signal

Cell

Cell

Cell

Cancer statistics, 2014

CA Cancer J. Clin

Adjuvant chemotherapy for non-small-cell lung cancer: the end of the beginning

J. Natl. Cancer Inst

Adjuvant chemotherapy in completely resected non-small-cell lung cancer

J. Clin. Oncol

Adjuvant chemotherapy for resectable non-small-cell lung cancer: where is it going?

Ann. Oncol

The mechanism of necroptosis in normal and cancer cells

Cancer Biol. Ther

The role of necroptosis, an alternative form of cell death, in cancer therapy

Expert Rev. Anticancer Ther

Clinical significance of phosphorylated Caspase-8 by Src in non-small cell lung cancer

NanFang Med J

HOXA13 promotes cancer cell growth and predicts poor survival of patients with esophageal squamous cell carcinoma

Cancer Res

RIPK1 both positively and negatively regulates RIPK3 oligomerization and necroptosis

Cell Death Differ

Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury

Nat. Chem. Biol

Original Articles
Dasatinib promotes paclitaxel-induced necroptosis in lung adenocarcinoma with phosphorylated caspase-8 by c-Src