Expression and prognostic significance of centromere protein A in human lung adenocarcinoma

doi:10.1016/j.lungcan.2012.04.007

Lung Cancer

Volume 77, Issue 2, August 2012, Pages 407-414

https://doi.org/10.1016/j.lungcan.2012.04.007 Get rights and content

Abstract

Background

Centromere protein A (CENP-A), one of the fundamental components of the human active kinetochore, is frequently upregulated in many cancers and plays important roles in cell cycle regulation, cell survival, and genetic stability. The aim of the present study was to explore the expression and prognostic significance of CENP-A in lung adenocarcinoma.

Experimental design

The expression of CENP-A was detected in 20 fresh human lung adenocarcinoma specimens and corresponding non-tumorous lung tissues by real-time polymerase chain reaction (RT-PCR) and Western blotting analysis. Using immunohistochemistry, we analyzed CENP-A protein expression in additional 309 lung adenocarcinomas. The clinicopathological and prognostic significance of CENP-A expression was analyzed.

Results

RT-PCR and Western blotting analysis revealed an enhanced expression of CENP-A in lung adenocarcinomas relative to adjacent non-tumorous lung tissues at both transcriptional and translational levels. Immunohistochemistry showed that 146 of 309 lung adenocarcinomas (47.3%) had high expression of CENP-A. CENP-A overexpression was significantly correlated with pathological grade (P = 0.009), pT status (P = 0.017), pN status (P = 0.002), pleural invasion (P = 0.013), high Ki-67 expression (P = 0.003), and P53 positivity (P = 0.001). Patients with high CENP-A expression had shorter overall survival time compared with those with low CENP-A expression. Multivariate analysis identified CENP-A as an independent prognostic factor for lung adenocarcinoma.

Conclusion

Our results demonstrate that elevated CENP-A expression is closely associated with lung adenocarcinoma progression and has an independent prognostic value in predicting overall survival for patients with lung adenocarcinoma.

Introduction

Lung cancer is one of the most common malignancies worldwide, causing about 1.4 million global deaths in 2008 [1]. Non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancers [2] and can be sub-divided into three histological subtypes (i.e. adenocarcinoma, squamous cell carcinoma, and large cell carcinoma) [3]. In the past 20 years, lung adenocarcinoma (∼40%) has become the most frequent subtype of NSCLC in most countries [4], [5]. To date, no standard prognostic marker has been established for NSCLC, especially lung adenocarcinoma.

Chromosome mis-segregation during mitosis is believed to be the main cause of aneuploidy, a hallmark feature of cancer cells that has been closely linked to tumorigenesis and cancer progression [6]. The kinetochore, which consists of centromeric DNA and associated proteins, plays a critical role in chromosome segregation and cell division [7]. Centromere protein (CENP)-A, a centromere-specific 17-kDa protein, is one of the first identified kinetochore components in humans. It is a unique histone H3-like protein only found in active centromeres and implicated in the epigenetic maintenance of centromere identity [8]. CENP-A plays important roles in cell cycle regulation and cell survival [9]. It has been documented that CENP-A expression is frequently elevated in numerous types of human cancers including hepatocellular carcinoma (HCC) and colorectal cancer [9], [10]. Tomonaga et al. [10] has provided evidence for a link between CENP-A overexpression and aneuploidy in colorectal cancers. Another member of the centromere protein family, CENP-H, has been found to be upregulated in NSCLC and predicts poor outcome in patients with this disease [11]. However, relatively little is known about the expression and clinical significance of CENP-A in lung adenocarcinoma.

In this study, we therefore assessed the expression of CENP-A in a series of lung adenocarcinoma specimens using immunohistochemistry, and investigated its associations with clinicopathologic parameters and overall survival in patients with lung adenocarcinoma receiving curative surgical resection.

Section snippets

Human tissues samples

Tumor samples from resection specimens were collected from two consecutive cohorts of patients with primary lung adenocarcinomas. Cohort A consisted of 20 patients, from whom fresh tumor samples coupled with adjacent nontumorous lung tissues 5–10 cm remote from the tumor for analysis of CENP-A expression by semi-quantitative and quantitative PCR and Western blot analyses as described below. All of the excised tissues were placed immediately in liquid nitrogen and stored at −80 °C. Cohort B

Overexpression of CENP-A in human lung adenocarcinoma tumor tissues

We examined the expression of CENP-A transcript using the qPCR assay in 20 pairs of snap-frozen lung adenocarcinoma and adjacent nonmalignant lung tissues. Lung adenocarcinoma specimens expressed a significantly higher level of CENP-A mRNA than did the corresponding nontumorous tissues (P < 0.01; Fig. 1A). Western blotting analysis further revealed that the CENP-A protein level was significantly higher in lung adenocarcinoma specimens compared to the corresponding nontumorous tissues (P < 0.01;

Discussion

In this report, we presented the first evidence that CENP-A mRNA transcript and protein levels were consistently increased in lung adenocarcinoma tissues relative to those in nontumorous lung tissues. Furthermore, we showed that the CENP-A expression determined by immunohistochemistry was significantly correlated with histological grade, pT status, pN status, pleural invasion status, Ki-67 labeling index, and P53 positivity. Most importantly, patients with high CENP-A expression had

Conflict of interest statement

None.

Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81072020 and No. 81172311) to S.H. Zhang.

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View full text

Expression and prognostic significance of centromere protein A in human lung adenocarcinoma

Abstract

Background

Experimental design

Results

Conclusion

Introduction

Section snippets

Human tissues samples

Overexpression of CENP-A in human lung adenocarcinoma tumor tissues

Discussion

Conflict of interest statement

Acknowledgments

Lung Cancer

Lung Cancer

Mod Pathol

Semin Cell Dev Biol

Curr Opin Cell Biol

Mol Cell

Methods

Semin Cell Dev Biol

Cancer Lett

J Thorac Oncol

Global cancer statistics

CA Cancer J Clin

Lung cancer

N Engl J Med

ShRNA-targeted centromere protein A inhibits hepatocellular carcinoma growth

PLoS One

Overexpression and mistargeting of centromere protein-A in human primary colorectal cancer

Cancer Res

Centromere protein H is a novel prognostic marker for human nonsmall cell lung cancer progression and overall patient survival

Cancer