In vivo and in vitro cell based model of lung adenocarcinoma from patients with pleural effusion

Abstract

Lung adenocarcinoma (LAC) is a common and aggressive form of lung cancer that is increasing in incidence among never smokers at younger age. Current treatment of patients with LAC is insufficient and there is a need for identification of effective biomarkers and development of therapy targets. These demands require also improved models for in vivo and in vitro experimentation. In this study, we describe the establishment of two LAC cell lines, termed LuCa-3 and LuCa-6. Both were derived from pleural effusion (PE) cells of LAC patients (L3 and L6) and both are readily propagated as tumor xenografts in immune-deficient mice. PE cells from the patient L6 exhibited also capacity for in vitro growth and were cultured in two forms: (i) as a suspension growing cell population, termed LuCa-6S, composed of non-clumping single cells; and (ii) as a monolayer-like culture, labeled LuCa-6A, exhibiting tight cell-to-cell and to culture surface adherence. Unique features of these two sublines and their cell clones are the capacity to convert from a non-clumping single-cell suspension into the adherent growth pattern and vice versa. Immunostaining of patients' tumor tissue xenografts and cultured subline cells displayed markers specific for the phenotype of human LAC. LuCa-6S and LuCa-6A cells did not reveal a noticeable disparity in quantitative growth characteristics. However, a number of differences were detected between these two cell populations manifested in detection or intensities of antigen expressions on cell surface (CD133, SFTPC) and in the nucleus (TTF-1) including pluripotent (OCT-4, SOX-2, NANOG) genes in cancer stem-like cells (CSCs). Dissimilarities between these two sublines were also detected in N-glycan profiles and in the sensitivity to natural killer cells. Salient features of these subline cell populations are responsiveness to selective upregulation of the pluripotent genes in subsets of CSCs via conversion of their growth patterns and/or by using culture stem media with growth factors. The described in vivo/in vitro model enables broader experimental approaches in studies of lung adenocarcinoma.

Differentiation of Sialyl Linkages Using a Combination of Alkyl Esterification and Phenylhydrazine Derivatization: Application for N-Glycan Profiling in the Sera of Patients with Lung Cancer

“…The most commonly occurring SAs in mammalians are N -acetylneuraminic acid (Neu5Ac) and N- glycolylneuraminic acid (Neu5Gc). Alterations in N -glycans including changes in the type of SA and their attachments have been associated with different diseases. − This knowledge points to the importance of a deeper investigation of glycosylation, that is, discrimination of the structural heterogeneity of glycans, their branching patterns, or linkage types …”