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Expression of ezrin correlates with malignant phenotype of lung cancer, and in vitro knockdown of ezrin reverses the aggressive biological behavior of lung cancer cells

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Tumor Biology

Abstract

Ezrin, one of the ezrin–radixin–moesin proteins, is involved in the formation of cell membrane processes such as lamellipodia and filopodia and acts as a membrane–cytoskeleton linker. Its aberrant expression correlates with development and progression of several human cancers. However, the expression of ezrin and its role in lung cancer are currently unknown. In this study, we performed ezrin small interfering RNA transfection in two lung cancer cell lines and examined the effects on malignant phenotypes in cancer cells by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, wound healing, and chamber transwell assays. Ezrin knockdown significantly reduced the proliferation, migration, and invasion of lung cancer cells in vitro. To address the possible mechanisms, we evaluated the expression of adhesion molecules E-cadherin and β-catenin by Western blot and reverse transcriptase-polymerase chain reaction analyses. The results demonstrated that downregulation of ezrin reduced β-catenin and increased E-cadherin at the protein level but had no effects on their mRNA levels, suggesting posttranscriptional regulation of these two adhesion molecules. Immunofluorescence assays revealed that ezrin knockdown restored membranous expression of E-cadherin and decreased cytoplasmic β-catenin in lung cancer cells. In addition, ezrin expression was immunohistochemically evaluated on 135 normal and 183 lung cancer tissues. The expression of ezrin was significantly higher in cancer samples than paired autologous normal lung tissues. In normal bronchial epithelium, ezrin was mainly localized on the apical membrane, while in lung cancers and metastatic foci, ezrin was primarily distributed in cytoplasm. Among lung cancer tissues, expression of ezrin was higher in the invasive front of primary lesions and the highest in lymphatic metastasis. Statistical analysis demonstrated that ezrin expression correlated significantly with lymphatic metastasis and advanced TNM stage. Our data suggest that ezrin may play a crucial role in governing the biological behavior of lung cancer.

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Abbreviations

BSA:

Bovine serum albumin

ERM:

Ezrin–radixin–moesin

ICAM:

Intercellular adhesion molecule

siRNA:

Small interfering RNA

DMSO:

Dimethyl sulfoxide

DAB:

Diaminobenzidine

PVDF:

Polyvinylidene fluoride

HRP:

Horseradish peroxidase

EB:

Ethidium bromide

NSCLC:

Non-small cell lung cancer

FCS:

Fetal calf serum

FITC:

Fluorescein isothiocyanate

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Financial support

This study received financial support from the National Natural Science Foundation of China (grant no. 30700806 to Q-C Li).

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Authors and Affiliations

  1. Department of Pathology, The First Affiliated Hospital and College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China

    Qingchang Li, Hui Gao, Hongtao Xu, Xin Wang, Yongqi Pan, Fengxia Hao, Xueshan Qiu & Enhua Wang

  2. Institute of Pathology and Pathophysiology, China Medical University, Shenyang, 110001, China

    Qingchang Li, Hui Gao, Hongtao Xu, Xin Wang, Yongqi Pan, Fengxia Hao, Xueshan Qiu & Enhua Wang

  3. Department of Pathology, Duke University Medical Center, Durham, NC, 27710, USA

    Maggie Stoecker & Endi Wang

Authors
  1. Qingchang Li
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  2. Hui Gao
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  3. Hongtao Xu
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  4. Xin Wang
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  9. Endi Wang
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  10. Enhua Wang
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Correspondence to Enhua Wang.

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Li, Q., Gao, H., Xu, H. et al. Expression of ezrin correlates with malignant phenotype of lung cancer, and in vitro knockdown of ezrin reverses the aggressive biological behavior of lung cancer cells. Tumor Biol. 33, 1493–1504 (2012). https://doi.org/10.1007/s13277-012-0400-9

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  • DOI: https://doi.org/10.1007/s13277-012-0400-9

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