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Mast cells (MCs) are part of tumor microenvironment, but the effect of hypoxia on the production of MC-derived cytokines has not been fully described. Using the hypoxia marker pimonidazole in vivo, we found that MCs were largely located in the low-oxygen areas within B16-F1 mice melanoma tumors. In vitro, hypoxia promoted ROS production, a ROS-dependent increase of intracellular calcium, and the production of MCP 1 (CCL-2) in murine bone marrow\u2013derived MCs. Hypoxia-induced CCL-2 production was sensitive to the antioxidant trolox and to nifedipine, a blocker of L-type voltage-dependent Ca2+ channels (LVDCCs). Simultaneously with CCL-2 production, hypoxia caused the ROS-dependent glutathionylation and membrane translocation of the \u03b11c subunit of Cav1.2 LVDCCs. Relationship between ROS production, calcium rise, and CCL-2 synthesis was also observed when cells were treated with H2O2. In vivo, high CCL-2 production was detected on hypoxic zones of melanoma tumors (where tryptase-positive MCs were also found). Pimonidazole and CCL-2 positive staining diminished when B16-F1 cell\u2013inoculated animals were treated with trolox, nifedipine, or the adenosine receptor 2A antagonist KW6002. Our results show that MCs are located preferentially in hypoxic zones of melanoma tumors, hypoxia-induced CCL-2 production in MCs requires calcium rise mediated by glutathionylation and membrane translocation of LVDCCs, and this mechanism of CCL-2 synthesis seems to operate in other cells inside melanoma tumors, with the participation of the adenosine receptor 2A.<\/jats:p>","DOI":"10.4049\/jimmunol.1801430","type":"journal-article","created":{"date-parts":[[2020,2,4]],"date-time":"2020-02-04T02:25:26Z","timestamp":1580783126000},"page":"1056-1068","source":"Crossref","is-referenced-by-count":35,"title":["Mast Cells Localize in Hypoxic Zones of Tumors and Secrete CCL-2 under Hypoxia through Activation of L-Type Calcium Channels"],"prefix":"10.1093","volume":"204","author":[{"given":"Itzel G","family":"Ram\u00edrez-Moreno","sequence":"first","affiliation":[{"name":"Departamento de Farmacobiolog\u00eda, Centro de Investigaci\u00f3n y de Estudios Avanzados (Cinvestav) , Tlalpan, 14330 Mexico City ,","place":["Mexico"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alfredo","family":"Ibarra-S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Departamento de Farmacobiolog\u00eda, Centro de Investigaci\u00f3n y de Estudios Avanzados (Cinvestav) , Tlalpan, 14330 Mexico City ,","place":["Mexico"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3278-2348","authenticated-orcid":false,"given":"Jorge Ivan","family":"Castillo-Arellano","sequence":"additional","affiliation":[{"name":"Instituto de Qu\u00edmica, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Ciudad Universitaria , 04510 Mexico City ,","place":["Mexico"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2661-2983","authenticated-orcid":false,"given":"Ulrich","family":"Blank","sequence":"additional","affiliation":[{"name":"Inserm U1149, CNRS ERL 8252, Universit\u00e9 Paris Diderot, Sorbonne Paris Cit\u00e9, Facult\u00e9 de M\u00e9decine, Site X. bichat, Laboratorie d'excellence INFLAMEX , 75018 Paris ,","place":["France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7332-3829","authenticated-orcid":false,"given":"Claudia","family":"Gonz\u00e1lez-Espinosa","sequence":"additional","affiliation":[{"name":"Departamento de Farmacobiolog\u00eda, Centro de Investigaci\u00f3n y de Estudios Avanzados (Cinvestav) , Tlalpan, 14330 Mexico City ,","place":["Mexico"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,2,15]]},"reference":[{"key":"2025030417510688200_r1","doi-asserted-by":"crossref","first-page":"648","DOI":"10.1016\/j.it.2017.04.001","article-title":"Rheostatic functions of mast cells in the control of innate and adaptive immune responses","volume":"38","author":"Frossi","year":"2017","journal-title":"Trends Immunol."},{"key":"2025030417510688200_r2","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1111\/imr.12629","article-title":"Non-IgE mediated mast cell activation","volume":"282","author":"Redegeld","year":"2018","journal-title":"Immunol. 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