Galectin-12 colocalizes with splicing factor-rich speckles and shuttles between the nucleus and cytoplasm in colon cancer cells

Eva Katzenmaier, Hans-Jürgen Stark, Johannes Gebert, Jürgen Kopitz

Abstract


Several members of the glycan-binding family of galectins have been linked to colon cancer initiation and progression while the role of galectin-12 in this malignancy is largely unexplored. In previous studies we observed expression of galectin-12 in normal colon epithelium in contrast to its lack of expression in colorectal cancer tissues. In order to gain insight into its molecular function we established a genetically engineered colon cancer model cell line, which facilitates inducible and reconstituted expression of LGALS12 transgene at physiological levels in an isogenic background. Regulation of transgene expression by doxycycline was confirmed at the transcript- and protein level in a time- and dose-dependent manner for two independent clones. Reconstituted galectin-12 expression did not affect cell morphology and proliferation. Analysis of its subcellular distribution showed that galectin-12 resides in and shuttles between the nucleus and cytoplasm. More detailed analysis by double-immunofluorescence and confocal microscopy revealed colocalization of galectin-12 with splicing-factor rich nuclear speckles, hinting towards a potential role of galectin-12 in pre-mRNA splicing and processing. Therefore, the established model cell line represents a powerful tool to investigate the functional impact of galectin-12 reconstitution on colon cancer tumorigenesis.


Keywords


colorectal cancer; lectins; galectins; nucleocytoplasmic shuttling; Tet-On system; Tet-regulated gene expression

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