Heterogeneity of microvesicles from cancer cell lines under inflammatory stimulation with TNF‐α

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.

Ambrisentan, an endothelin receptor type A-selective antagonist, inhibits cancer cell migration, invasion, and metastasis

“…S2 ). They were cultured in 10% panexin NTA Serum (Pan-Biotech, Aidenbach, Germany) or 10% fetal bovine serum (FBS) + DMEM high glucose medium (Gibco-ThermoFisher Scientific, Waltham, MA, USA) as previously described 81 . The mouse mammary gland tumor cell line 4T1 36 was generously provided by Dr Jo Van Ginderachter (Vrije Universiteit Brussel, Belgium).…”

“…The migration of cancer adherent cell lines (COLO357 and OvCar3) was analyzed using the Oris migration assay 81 . COLO357 cells were cultured and allowed to migrate into the free space in the middle of the well for 48 h in the presence or absence of 100 µM PAR2 agonistic peptide (MoBiTec GmbH, Göttingen, Germany) and/or Ambrisentan.…”

Modeling of the immune response in the pathogenesis of solid tumors and its prognostic significance

“…MV release is also dependent on cytokines. Incubation of cell lines derived from various tumors with TNF-α has been found to enhance the release of MVs [324]. Leukemia cell-derived MVs have also been found to disrupt lymphocyte functions by inducing T cell exhaustion [325].…”

The role of microvesicles and its active molecules in regulating cellular biology

“…In addition, the pro‐coagulant cascade and mediating cartilage regeneration are also important physiological functions of active lipids. Phosphatidylserine and tissue factor (TF), which are expressed by MVs, are directly involved in the activation of the coagulation cascade in the body . TF is inhibited in resting cells.…”