Metastatic melanoma is one of the most aggressive forms of skin cancer and has a poor prognosis. We have previously identified Annexin A1 (ANXA1) as a potential murine melanoma-spreading factor that may modulate cell invasion by binding to formyl peptide receptors (FPRs). Here, we report that (1) in a B16Bl6 spontaneous metastasis model, a siRNA-induced decrease in tumoral ANXA1 expression significantly reduced tumoral MMP2 activity and number of lung metastases; (2) in a retrospective study of 61 patients, metastasis-free survival was inversely related to ANXA1 expression levels in primary tumors (HR 3.15 [1.03-9.69], p = 0.045); (3) in human melanoma cell lines, ANXA1 level was positively correlated with in vitro invasion capacity whereas normal melanocytes contained low ANXA1 levels, and (4) the ANXA1 N-terminal peptide ANXA12-26 stimulated MMP2 activity after interaction with FPRs and significantly stimulated the in vitro invasion of melanomas by acting on FPRs. These findings identify ANXA1 as a proinvasive protein in melanoma that holds promise as a potential prognostic marker and therapeutic target.

The cyclization of anticancer drugs into active intermediates has been reported mainly for DNA alkylating molecules including nitrosoureas. We previously defined the original cytotoxic mechanism of anticancerous N-phenyl-N'-(2-chloroethyl)ureas (CEUs) that involves their reactivity towards cellular proteins and not against DNA; two CEU subsets have been shown to alkylate β-tubulin and prohibitin leading to inhibition of cell proliferation by G₂/M or G₁/S cell cycle arrest. In this study, we demonstrated that cyclic derivatives of CEUs, N-phenyl-4,5-dihydrooxazol-2-amines (Oxas) are two- to threefold more active than CEUs and share the same cytotoxic properties in B16F0 melanoma cells. Moreover, the CEU original covalent binding by an ester linkage on β-tubulin Glu198 and prohibitin Asp40 was maintained with Oxas. Surprisingly, we observed that Oxas were spontaneously formed from CEUs in the cell culture medium and were also detected within the cells. Our results suggest that the intramolecular cyclization of CEUs leads to active Oxas that should then be considered as the key intermediates for protein alkylation. These results will be useful for the design of new prodrugs for cancer chemotherapy.

with systemic spread of the disease. Beside the destructive treatments, the intralesional BCG application (IBA) is a method of choice with initial CRR up to 80%. The BCG-induced immune reaction is biphasicimmediate humoral (pro-apoptotic) and delayed cellmediated (cytotoxic).Methods and objects: A novel method of treatment of ITM is introduced. Approximately 6 h. after the conventional IBA (with the first appearance of the clinical symptoms of fever, malaise, muscle and joint pains) electro-poration (EP) is performed to enhance the entry of the released pro-apoptotic cytokines into the tumor cells. During 2009-2011, 17 pts with 287 (6-43 per pt) ITM were treated, 4 with BCG alone (83 lesions) and 13 -BCG + EP (204 lesions).Results: Lesions less than 2 mm were with CR for both methods (32 and 66 lesions respectively). Lesions 2-5 mm were with 32% PR for the BCG group (12 from 37 lesions) and re-application was needed while for the BCG + EP group the PR was only 7% (6 from 82 lesions). Lesions > 5 mm were with 57% PR for the BCG group (8 from 14 lesions) and re-application was needed while for the BCG + EP group the PR was only 11% (6 from 56 lesions). Discussion:The electro-immunotherapy is more effective than the IBA alone for superficial ITM larger than 2 mm. We prefer IBA to the destructive treatment of the superficial ITM because the lower ratio of consequent regional lymphnode mets and we presume that a better loco-regional control of the disease could be achieved. The systemic spread of the disease seems to be related rather to the appearance of deep ITM and even after their successful treatment by surgery or isolated limb perfusion the final prognosis remains unchanged.