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N A van Larebeke

Publications and source records attributed to N A van Larebeke.

3 recordsLinked to original sources

An anti-invasive concentration of the alkyl-lysophospholipid ET-18-OCH3 enhances the motility of embryonal chick heart cells cultured on solid substrate.

Pretreatment of embryonal chick heart fragments with ET-18-OCH3 is known to induce resistance to invasion by several malignant cell lines. Embryonal chick heart fragments or cell suspensions prepared from such fragments were explanted on solid substrate and treated in medium with 10 micrograms/ml ET-18-OCH3 or with drug-free medium (control) for 48 h. This medium was washed away and replaced by drug-free fresh medium. Twenty-four to 48 h later the fast plasma membrane movements (involved in ruffling, blebbing, fast shape change and fast translocation) were quantified using a simple method based on subtracting two video images taken with an interval of 28 s. The ET-18-OCH3-treated cells showed a higher intensity of fast plasma membrane movements than control cells. Cells around a treated explant did not show the same radial alignment as in controls, suggesting loss of contact inhibition of movement. Cells from a cell suspension derived from a treated fragment showed faster translocation on solid substrate and faster shape change. We speculate that increased motility of host cells may be involved in resistance to invasion.

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Simple method for quantification of fast plasma membrane movements.

We present a method for the quantification of the fast plasma membrane movements that are involved in ruffling, blebbing, fast shape change, and fast translocation. The method is based on the Kontron Vidas image analysis computer program. Video images from cells viewed through an inverted microscope were transmitted to the computer. The procedure was as follows: 4 consecutive video images were averaged (image 1); 28 s later a second set of 4 video images was averaged (image 2); image 2 was subtracted from image 1 and the grey level of each pixel of the resulting image was increased with 128 grey level units, resulting in the subtraction image, showing a uniform grey background speckled with brighter and darker spots corresponding to areas of movement. These spots were discriminated and turned into white objects against a black background. Interactive editing was used to delete artefacts that resulted from floating debris. The total area of the discriminated objects was measured, and the parameter motile area in micron2 per cell was calculated. We have applied our method to the study of motility induced in epithelial cell lines by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate and by epidermal growth factor.

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Invasive epithelial cells show more fast plasma membrane movements than related or parental non-invasive cells.

Fast plasma membrane movements (FPMM) are involved in ruffling, blebbing, fast shape change, and fast translocation. A simple method for the quantification of FPMM was used to study the relation between FPMM and invasive capacity in five pairs of invasive and noninvasive variants from four different epithelial cell types. The human mammary cell line MCF-7/6, the ras-transformed dog kidney cell line ras-MDCK, the ras-transformed mouse mammary gland cell lines NM9-ras-12 and NM-f-ras-TD, and spontaneously transformed late passage mouse lens explant MLE cells, all of which were invasive in vitro, showed more FPMM in our measurements and displayed more ruffling activity on time-lapse video films than the related or parental MCF-7/AZ, MDCK-3, NM9, and NM-f cell lines and early passage MLE cells, none of which were invasive. Interestingly, induction of invasive capacity in MCF-7/AZ cells by retinoic acid was accompanied by an increase in FPMM, but speed of translocation was not increased. Together these observations support the hypothesis that a certain level of FPMM is a prerequisite for invasive capacity.

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