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Heterogeneity of microvesicles from cancer cell lines under inflammatory stimulation with TNF-α
Frank Gieseler1, Corinna Plattfaut1, Tabea Quecke1
1Section Experimental Oncology, University Hospital and Medical School (UKSH), University of Luebeck, Luebeck, 23538, Germany.
Abstract:
Microvesicles (MVs) represent a subgroup of extracellular vesicles (EVs) emerging from various cells by blebbing of their outer membrane. Therefore, they share features such as membrane composition and antigenicity with their parental cells. Released by many immune and tumor cells, MVs act as intercellular messengers, account for horizontal gene transfer and can activate the coagulation system. With the aim to investigate their relevance for tumor cell biology, we characterized MVs released by human tumor cell lines of various origins in the absence or presence of TNF-α. After stimulation, we used the combination of low and high-speed centrifugation to enrich MVs from cell culture supernatants. We analyzed the presentation of phosphatidylserine (PS) and tissue factor (TF) activity on the cell surface and investigated their potency to induce tumor cell migration. In all tumor cell lines, TNF-α stimulation enhanced the release of MVs. While the expression of PS was universally increased, an elevated activity of procoagulant TF could be detected on MVs from lung, pancreatic, and colon carcinoma, but not from breast and ovarian cancer cell lines. Functionally, TNF-α stimulation significantly increased the potency of MVs to induce tumor cell migration. In conclusion, inflammatory conditions promote the release of MVs with increased procoagulant activity from tumor cell lines in vitro. PS-containing and TF-expressing MVs may account for systemic activation of the coagulation system as seen in cancer patients and, since they induce tumor cell migration, they may serve as biomarkers for tumor progression.
Insights
Tumor cells release more microvesicles (MVs) when exposed to inflammation. These MVs, carrying phosphatidylserine (PS) and tissue factor (TF), promote tumor cell migration and may indicate cancer progression.
Area of Science:
- Extracellular vesicle research
- Cancer biology
- Immunology
Background:
- Microvesicles (MVs) are extracellular vesicles (EVs) released from the cell membrane, sharing characteristics with parent cells.
- MVs from immune and tumor cells act as messengers, mediate gene transfer, and can activate coagulation.
- Tumor-derived MVs play roles in cancer progression and may be linked to coagulation abnormalities in cancer patients.
Purpose of the Study:
- To investigate the characteristics and functions of MVs released by human tumor cell lines under inflammatory conditions (TNF-α stimulation).
- To analyze the expression of phosphatidylserine (PS) and tissue factor (TF) activity on MVs.
- To assess the impact of MVs on tumor cell migration.
Main Methods:
- Human tumor cell lines were cultured with or without TNF-α stimulation.
- MVs were enriched from cell culture supernatants using differential centrifugation.
- PS exposure and TF activity on MVs were analyzed.
- The effect of MVs on tumor cell migration was evaluated.
Main Results:
- TNF-α stimulation significantly increased MV release from all tested tumor cell lines.
- PS expression was universally elevated on MVs after TNF-α stimulation.
- Procoagulant TF activity was detected on MVs from lung, pancreatic, and colon cancer cell lines, but not breast or ovarian.
- TNF-α-stimulated MVs demonstrated enhanced potency in inducing tumor cell migration.
Conclusions:
- Inflammatory conditions enhance the release of MVs with increased procoagulant activity from tumor cells in vitro.
- PS- and TF-expressing MVs may contribute to systemic coagulation activation in cancer patients.
- These MVs could serve as potential biomarkers for tumor progression due to their role in migration.
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