Identification and Characterization of Different Subpopulations in a Human Lung Adenocarcinoma Cell Line (A549)

Maria Virginia CROCE1, Andrea Gladys COLUSSI1, Mike R PRICE2, Amada SEGAL-EIRAS1

1Centro de Investigaciones Inmunológicas Básicas y Aplicadas (CINIBA), Universidad Nacional de la Plata, La Plata, Argentina
2Cancer Research Laboratories, School of Pharmaceutical Sciences, University of Nottingham, Nottingham, UK


The morphology, cell growth, antigenic expression and tumorigenicity of cell subpopulations from the A549 lung adenocarcinoma isolated by Percoll gradient separation have been analysed. Four subpopulations were obtained (subpopulations A, B, C and D). Immunocytochemical analysis of several antigens was performed with monoclonal antibodies (MAbs): MUC1 mucin (C595, HMFG1 and HMFG2), MUC5B (PANH2); gp230 (PANH4); carbohydrate antigens including sialyl Lewis x (KM93), Tn antigen (83D4), Lewis y (C14); 5, 6, 8, 17 and 19 cytokeratins and p53. The cell population D tended to form cell aggregates that piled up on the monolayer similar to overgrowth cultures of the A549 parental cell line, whereas A, B and C cell subpopulations formed well spread monolayers. Both parental A549 and subpopulation D secreted abundant mucus. The topographic distribution and secretion production were correlated with tumorigenic assays since only subpopulation D grew in nude mice exhibiting reduced latency period; these characteristics correlated with the fast growth of the subpopulation D in vitro. Immunocytochemical analysis demonstrated that subpopulation D showed greater expression of MUC1 mucin and carbohydrate antigens such as Tn antigen, sialyl Lewis x and Lewis y and less expression of cytokeratins, p53, MUC5B and gp230; conversely, subpopulations A, B and C showed the opposite antigenic profile. Our results illustrate heterogeneity in the A549 cell line; subpopulations A, B and C retained characteristics of more differentiated adenocarcinoma while subpopulation D displayed features of a less differentiated tumor line. Pathology & Oncology Research, Vol 5, Nr 3, 197-204, 1999

Key words: A549; lung adenocarcinoma; cell subpopulations

Received: Jun 30, 1999; accepted: Jul 21, 1999
Correspondence: Amada SEGAL-EIRAS, Centro de Investigaciones Inmunológicas Básicas y Aplicadas (CINIBA), Universidad Nacional de la Plata, Calle 60 y 120 La Plata , Argentina; Tel: 54 2214214756 / 342, Fax: 54 221 4259115 / 6; E-mail:

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