Effect of cigarette smoke on the mRNA and protein expression of carcinoembryonic antigen (CEA), a possible chemoattractant for neutrophils in human bronchioloalveolar tissues.
BACKGROUND
The concentration of carcinoembryonic antigen (CEA), known as a marker of malignant transformation and chronic inflammation, is increased in bronchoalveolar lavage fluid obtained from smokers compared with fluid from non-smokers. This study investigated the mechanism and biological significance of CEA production in the lungs of smokers by evaluating protein and mRNA expression in non-carcinomatous lung parenchymal tissues and in cell lines derived from human fetal lung.


METHODS
Lung parenchymal tissue free from cancer or an inflammatory lesion was obtained from five non-smokers (four with lung cancer, one with pulmonary mycetoma), five ex-smokers (all with lung cancer except for one with mesothelioma), and 14 smokers (nine with lung cancer, five with emphysema) at surgery or necropsy. Cancer tissue was also collected simultaneously from the subjects with lung cancer. CEA protein in the tissue homogenates was measured by enzyme linked immunoassay. CEA mRNA expression in the non-carcinomatous parenchymal tissue and cancer tissue was evaluated by in situ hybridisation using CEA specific riboprobe and was semiquantitated by counting the number of silver grains per cell. CEA mRNA expression was also compared in three cell lines derived from human fetal lung (IMR-90, MRC-9, and CCD-14Br) after in vitro stimulation with medium exposed to cigarette smoke or air. Chemoattractant activity of purified CEA for neutrophils and monocytes was also studied in vitro.


RESULTS
CEA content in non-carcinomatous lung tissue was increased in smokers with emphysema (mean (SD) 38.0 (9.2) ng/mg protein) or with lung cancer (38.2 (21.6)) compared with non-smokers (11.0 (5.4)) or ex-smokers (5.9 (2.2)). CEA mRNA expression in non-carcinomatous tissue, expressed by average number of grains per cell, was also increased in smokers with emphysema (mean (SD) 11.2 (4.1)) or with lung cancer (14.0 (8.4)) compared with non-smokers (3.1 (0.6)) or ex-smokers (4.0 (1.7)). CEA content in carcinomatous tissues was 42.8 (37.3) for non-smokers, 38.2 (42.4)) for ex-smokers, and 59.0 (22.5) for smokers. The CEA content in carcinomatous tissue was higher than in non-carcinomatous tissue, but there was no difference between non-smokers, ex-smokers, and smokers. The numbers of grains per cell in carcinomatous tissue were higher than in non-carcinomatous tissues, but not different among non-smokers (30.3 (3.9)), ex-smokers (38.3 (13.8)), and smokers (44.3 (5.2)). CEA mRNA expression in the cell lines was upregulated after the incubation with smoke-treated medium. Purified CEA was chemoattractant for neutrophils but not for monocytes in vitro.


CONCLUSIONS
mRNA and protein expression of CEA were increased in the normal lung tissue from smokers compared with non-smokers or ex-smokers. Since CEA content and mRNA expression were no different between smokers with non-small cell lung cancer and those with non-carcinomatous disease, it is unlikely that CEA expression in non-carcinomatous lung parenchymal tissue was influenced by the presence of the tumour and is consistent with the effect of smoking. This is supported by in vitro studies which show that cigarette smoke could induce CEA mRNA expression in fetal lung derived cells. In addition, CEA might play a part in recruitment of neutrophils into the lower respiratory tract.
Ohwada
A
A
Department of Respiratory Medicine, Juntendo University School of Medicine, Tokyo, Japan.
Takahashi
H
H
Nagaoka
I
I
Iwabuchi
K
K
Mikami
O
O
Kira
S
S
D000368:Aged; D001483:Base Sequence; D002272:Carcinoembryonic Antigen / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000502:physiology*; D017077:Culture Media, Conditioned; D005260:Female; D006801:Humans; D017403:In Situ Hybridization; D008168:Lung / Q000378:metabolism*; D008175:Lung Neoplasms / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009000:Monocytes / Q000502:physiology; D009504:Neutrophils / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012907:Smoking / Q000378:metabolism*
D002272:Carcinoembryonic Antigen; D017077:Culture Media, Conditioned; D012333:RNA, Messenger