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. 2008 Aug 25;114(4):275-83.
doi: 10.1002/cncr.23596.

"VSports手机版" Magnetic enrichment of bronchial epithelial cells from sputum for lung cancer diagnosis

Qi Qiu  1 , Nevins W Todd, Ruiyun Li, Hong Peng, Zhenqiu Liu, Harris G Yfantis, Ruth L Katz, Sanford A Stass, Feng Jiang
Affiliations

"V体育官网" Magnetic enrichment of bronchial epithelial cells from sputum for lung cancer diagnosis

"VSports注册入口" Qi Qiu et al. Cancer. .

Abstract

Background: Sputum is an easily accessible diagnostic material for lung cancer early detection by cytologic and molecular genetic analysis of exfoliated airway epithelial cells. However, the use of sputum is limited by its cellular heterogeneity, which includes >95% macrophages and neutrophils and only about 1% bronchial epithelial cells VSports手机版. We propose to obtain concentrated and purified bronchial epithelial cells to improve early detection of lung cancer in sputum samples. .

Methods: Sputum was collected from patients with stage I nonsmall-cell lung cancer, cancer-free smokers, and healthy nonsmokers. Magnetic-assisted cell sorting (MACS) with anti-CD14 and anti-CD16 antibody beads were used to enrich bronchial epithelial cells by depleting macrophages and neutrophils from sputum. Fluorescence in situ hybridization (FISH) analysis for detection of FHIT deletion and cytology were evaluated in the enriched specimens V体育安卓版. .

Results: The bronchial epithelial cells were concentrated to 40% purity from 1. 1% of the starting population, yielding an average of 36-fold enrichment and at least 2. 3 x 10(5) cells per sample. Detecting FHIT deletions for lung cancer diagnosis produced 58% sensitivity in the enriched sputum, whereas there was 42% sensitivity in the unenriched samples (P = . 02) V体育ios版. Cytologic examination of the enriched sputum resulted in 53% sensitivity, as compared with 39% sensitivity in unenriched sputum (P = . 03). Furthermore, only 2 cytocentrifuge slides of the unenriched sputum were needed for the analyses, as compared with up to 10 cytocentrifuge slides required from the unprocessed specimens. .

Conclusions: The enrichment of bronchial epithelial cells could improve the diagnostic value of sputum and the efficiency of genetic and cytologic analysis of lung cancer VSports最新版本. .

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Figures

FIGURE 1
FIGURE 1
Fluorescence in situ hybridization (FISH) analyses of normal peripheral blood lymphocytes. Normal interphase cells show 2 green signals from the FHIT probe and 2 red signals from the CEP3 probe. Original magnification, ×400.
FIGURE 2
FIGURE 2
Fluorescence in situ hybridization (FISH) analyses of exfoliated cells from sputum specimens. (A) A cytocentrifuge slide prepared from enriched sputum of a lung cancer patient shows a high percentage of bronchial epithelial cells characterized by elongated nuclei stained with DAPI. FISH analysis of the sample shows hemizygous deletion of FHIT represented by 1 green signal of FHIT probe and 2 signals of the CEP3 probe in some bronchial epithelial cells. Original magnification, ×400. (B) A cytocentrifuge slide prepared from unenriched sputum from the same patient shows a high percentage of nonepithelial cells. The nonepithelial cells are mainly neutrophils and macrophages, which have several round nuclei or nucleus that appear segmented and has lobes with weak signals of the probes. Original magnification, ×400.
FIGURE 3
FIGURE 3
Cytological analysis of sputum sample before and after enrichment. (A) A cytocentrifuge slide prepared from unenriched cells of 1 sputum sample shows that the majority of the cells are neutrophils and macrophages with abundant debris, whereas bronchial epithelial cells are scarce (Papanicolaou stain; original magnification, ×200). (B) A cytocentrifuge slide prepared from enriched cells from the same sample shows more than half of cells were bronchial epithelial cells with clear background (Papanicolaou stain; original magnification, ×200).
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