Membrane and Cytoplasmic Marker Exchange Between Malignant Neoplastic Cells and Fibroblasts Via Intermittent Con-tact: Increased Tumour Cell Diversity Independent of Genetic Change

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David, Manu; Huynh, Minh; Kelly, Elizabeth; Rizos, Helen; Coleman, Hedley; Rogers, Glynn; Zoellner, Hans


2012-06-01


Journal Article


Journal of Pathology


228


4


495 - 505


We earlier demonstrated human osteosarcoma cells (SAOS-2) induce contact-dependent apoptosis in endothelium, and expected similar apoptosis in Human Gingival Fibroblasts (h-GF) using SAOS-2 alkaline phosphatase (AP) to identify cells. However, h-GF apoptosis did not occur, despite reduction in AP-negative h-GF number (p0.01), and enhancement of this by h-GF TNF−α pre-treatment (p0.01). We suggest TNF−α-enhanced transfer of membrane AP from SAOS-2 to h-GF would explain these data. This idea was investigated using fluorescence pre-labeled cells and confocal laser scanning microscopy. Co-cultures of membrane labeled h-GF (marker-DiO) and SAOS-2 (marker-DiD) generated dual-labeled cells primarily at the expense of single labeled h-GF (p0.001), suggesting predominant membrane transfer from SAOS-2 to h-GF. However, opposite directional transfer predominated when membrane labels were reversed, and SAOS-2 further expressed green fluorescent protein (GFP) in cytoplasm and nuclei, and h-GF additionally bore nuclear label (Syto59) (p0.001). Cytoplasmic exchange was investigated using h-GF pre-labeled with cytoplasmic DDAO-SE and nuclear Syto59, co-cultured with SAOS-2 expressing GFP in cytoplasm and nuclei, and predominant cytoplasmic marker transferred from h-GF to SAOS-2 (p0.05). Pre-treating h-GF with TNF−α increased exchange of membrane markers (p0.04), but did not affect either cell surface area profile or circularity. Dual-labeled cells had morphological phenotype differing from SAOS-2 and h-GF (p0.001). Time-lapse microscopy revealed extensive migration of SAOS-2 and cell process contact with h-GF, with the appearance of SAOS-2 indulging in ‘cellular sipping’ from h-GF. Similar exchange of membrane was seen between h-GF and melanoma cell lines MeIRMu, NM39, WM175, NM200B12 and U205, as well as with ovarian carcinoma cells Pe01, Pe04 and Colo3/6, while cytoplasmic sharing was also seen in all cell lines other than U205 melanoma cells. We suggest that in some neoplasms, cellular sipping may contribute to pleomorphism and the generation of diverse tumor cell populations independent of genetic change, raising the possibility of a further as yet uninvestigated role in tumour progression.


Fibroblasts, Osteosarcoma Cells, Tumour Necrosis Factor-α, Membrane Exchange, Cytoplasmic Exchange


www.thejournalofpathology.com


nicta:5886


David, Manu; Huynh, Minh; Kelly, Elizabeth; Rizos, Helen; Coleman, Hedley; Rogers, Glynn; Zoellner, Hans. Membrane and Cytoplasmic Marker Exchange Between Malignant Neoplastic Cells and Fibroblasts Via Intermittent Con-tact: Increased Tumour Cell Diversity Independent of Genetic Change. Journal of Pathology. 2012-06-01; 228(4):495 - 505. <a href="http://hdl.handle.net/102.100.100/100581?index=1" target="_blank">http://hdl.handle.net/102.100.100/100581?index=1</a>



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