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首页> 外文期刊>Journal of Molecular Biochemistry >Galectin-12 colocalizes with splicing factor-rich speckles and shuttles between the nucleus and cytoplasm in colon cancer cells
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Galectin-12 colocalizes with splicing factor-rich speckles and shuttles between the nucleus and cytoplasm in colon cancer cells

机译：Galectin-12与富含剪接因子的斑点共定位，并在结肠癌细胞的细胞核与细胞质之间穿梭

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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.
机译：半乳凝素的聚糖结合家族的几个成员已与结肠癌的发生和发展有关，而半乳凝素12在这种恶性肿瘤中的作用尚待进一步研究。在以前的研究中，我们观察到了半乳糖凝集素12在正常结肠上皮中的表达，而在结肠直肠癌组织中缺乏表达。为了深入了解其分子功能，我们建立了基因工程结肠癌模型细胞系，该细胞系可促进在等基因背景中生理水平的诱导型和重组型LGALS12转基因表达。对于两个独立的克隆，以时间和剂量依赖性的方式在转录和蛋白质水平上证实了强力霉素对转基因表达的调节。重构的galectin-12表达不影响细胞形态和增殖。分析其亚细胞分布表明，galectin-12驻留在细胞核与细胞质之间，并在细胞核与细胞质之间穿梭。通过双重免疫荧光和共聚焦显微镜进行的更详细的分析显示，galectin-12与富含剪接因子的核斑点共定位，暗示了galectin-12在mRNA剪接和加工中的潜在作用。因此，已建立的模型细胞系代表了研究半乳凝素12重构对结肠癌肿瘤发生的功能影响的强大工具。

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