The induction of Maspin expression by a glucosamine-derivative has an antiproliferative activity in prostate cancer cell lines

Abstract

A B S T R A C TMammary serine protease inhibitor or Maspin has been characterized as a class II tumor suppressor gene in several cancer types, among them prostate cancer (CaP). Androgen ablation is an effective therapy for CaP, but with short-term effectiveness, thus new therapeutic strategies are actively sought. The present study is aimed to explore the effects of a glucosamine derivative, 2-(N-Carbobenzyloxy)L-phenylalanylamido-2-deoxy-β-D-glucose (NCPA), on two CaP cell lines, PC3 and LNCaP.In particular we analyzed the impact of NCPA on Maspin production, cell viability and cell cycle progression and apoptosis/necrosis pathway activation in PC3 and LNCaP cell lines.NCPA is able to stimulate Maspin production in PC3 and not in LNCaP cell lines. NCPA blocks the PC3 cell cycle in G1 phase, by inhibiting Cyclin D1 production and induces the apoptosis, therefore interfering with aggressiveness of this androgen-insensitive cell line. Moreover, NCPA is able to induce the expression of Maspin in LNCaP cell line treated with androgen receptor inhibitor, Bicalutamide, and in turn to stimulate the apoptosis of these cells.These findings suggest that NCPA, stimulating the endogenous production of a tumor suppressor protein, could be useful in the design of new therapeutic strategies for treatment of CaP. (M. Lopreiato), raffaellaguazzo@tiscali.it (R. Guazzo), desantim@ao-siena.toscana.it (M.M. de Santi), margherita.eufemi@uniroma1.it (M. Eufemi), roberto.scandurra@uniroma1.it (R. Scandurra), anna.scottodabusco@uniroma1.it (A. Scotto d'Abusco).

Split Gp41‐1 intein splicing as a model to evaluate the cellular location of the oncosuppressor Maspin in an in vitro model of osteosarcoma

“…Previously, it has been described that the nuclear localization of the Maspin is indicative of a favorable prognosis in the cancer progression 25,28 . Therefore, the nuclear/perinuclear localization of Maspin, in our experimental model, is a particularly intriguing result.…”

“…Previously, it has been described that the nuclear localization of the Maspin is indicative of a favorable prognosis in the cancer progression. 25,28 Therefore, the nuclear/perinuclear localization of Maspin, in our experimental model, is a particularly intriguing result. In this study, RFP fused-Maspin protein was obtained using the natural split gp41-1 inteins to analyze its cellular localization.…”

“…Previously, in our lab we found that the expression of Maspin, a class II tumor suppressor protein, was able to inhibit the invasiveness of osteosarcoma cells 27 and the cell cycle progression in two carcinoma prostate cell lines, 28 suggesting that the expression of this protein in cancer cells could be suggestive of a favorable prognosis. We decided to fuse the Maspin gene with the C‐construct, with the aim to follow the expression of this protein and its localization into the cells through the fluorescence produced by the reconstituted RPF protein as a result of IntN and IntC recombination.…”

“…Previously, we studied the efficacy of this protein to counteract the metastatic activity in osteosarcoma cell lines, finding that the expression of Maspin and its nuclear localization were able to decrease the β1 integrin expression in cell membrane and in turn affect the osteosarcoma cell invasiveness 27 . Moreover, we studied the effect of Maspin production in prostate carcinoma (CaP) cell lines, finding that its expression was able to block the cell cycle progression in PC3 line, which represent a hormone‐insensitive and later stage CaP, through the inhibition of Cyclin C1 expression 28 …”

“…27 Moreover, we studied the effect of Maspin production in prostate carcinoma (CaP) cell lines, finding that its expression was able to block the cell cycle progression in PC3 line, which represent a hormone-insensitive and later stage CaP, through the inhibition of Cyclin C1 expression. 28 The aim of the present manuscript, has been to follow the localization of Maspin in osteosarcoma cell lines, using two constructs containing the two moieties N-and C-split inteins and two moieties of Red Fluorescent Protein (RFP). In particular, to appreciate the recombination between IntN and IntC, through a model system in which two constructs, one containing IntN plus the N moiety of RFP and the second one containing IntC plus the C moiety of RFP plus the sequence coding for Maspin.…”