Effect of Increased Extracellular Ca⁺⁺ on Microvesicle Production and Tumor Spheroid Formation

Research on the composition of the tumor micro-environment has demonstrated that membrane delimited microvesicles are shed from many types of malignant tumors, in the peripheral blood of cancer patients as well as in culture media of tumor cells propagated in vitro (Ginestra et al. Anticancer Res 18:3433–3437, 1998). Their documented effects involve the activation of signal transduction pathways by cellular cross-talk that are associated with epigenetic mechanisms that may be important in tumor progression, metastasis, and the activation of angiogenesis (Distler et al. Arthritis Rheum 52:3337–3348, 2005). Live cell imaging microscopic studies conducted in our laboratory of the formation of solid tumor spheroids in vitro show that the shedding of microvesicular structures from tumor cells occurs during this process. The observed properties of the tumor microvesicles suggest a role in solid tumor formation and intercellular communication. The tumor associated microvesicles were shown to be non-apoptotic based on the absence of fluorescent nuclear staining by acridine orange/ethidium bromide staining. Increased concentration of extracellular Ca⁺⁺ [5–20 mM] resulted in an increase in the production of tumor-derived microvesicles and also to result in the formation of tumor spheroids whose size was considerably smaller than controls. Increased extracellular [Ca⁺⁺] was also observed to induce the rapid dissociation of solid tumor spheroids to smaller cell aggregates in the absence of significant apoptosis.