Preparation and characterization of differently aggregated colorectal carcinoma cell subpopulations from surgical...

The loss of intercellular adhesion within the primary tumor is one of the key events leading to metastasis. Although a number of adhesion molecules involved in intercellular adhesion have been described in experimental systems, the clinical relevance of many of these molecules still has to be determined. We tried to assess the contribution of membrane-bound carbohydrates and of E-Cadherin, CEA, and Sia-LeA for intercellular adhesion of cells isolated from colorectal carcinoma tissue directly obtained from the surgeon. A subpopulation of nonaggregating cells was prepared by means of slowly passing of freshly isolated cells through a series of sieves with decreasing mesh widths. Nonaggregating cells differed mainly in two aspects from aggregated cells: (i) determination of lectin binding and of specific sialytransferase activities revealed enhanced alpha2,6-sialylation of nonaggregating cells, and (ii) staining with specific antibodies documented a loss of E-Cadherin reactivity of such cells. An enhanced activity of beta-galactoside alpha2,6-sialytransferase (ST6Gal 1) was found in metastasizing colorectal carcinomas; however, its biological function has to be shown. Our results suggest that ST6Gal 1 is responsible for reduced homotypic aggregation of colorectal carcinoma cells and may thus facilitate the release of single cells from the primary tumor.