Claudio Tabolacci

Antitumor properties of aloe-emodin and induction of transglutaminase 2 activity in B16-F10 melanoma cells.

AIMS Aloe-emodin (AE), a natural hydroxyanthraquinone compound, has been reported as a potential anticancer agent. We studied the antineoplastic properties of AE on highly metastatic B16-F10 melanoma murine cells. MAIN METHODS Cell proliferation was assessed by cell counting and viability was investigated using MTT and Trypan Bleu exclusion tests. As a growth marker, we determined intracellular polyamine levels by high performance liquid chromatography. Then, we evaluated transglutaminase 2 (TG2) activity, protoporphyrin IX accumulation and melanin content as differentiative markers. Tyrosinase activity was checked by DOPA-staining assay. The antimetastatic effect of AE was evaluated by means of a series of in vitro metastatic assays, including aggregation, wound healing migration, adhesion, 3D-invasion, circular invasion and the Boyden chamber invasion assays. Gelatin zymography was performed to evaluate metalloproteinase activities. KEY FINDINGS Our results demonstrated inhibitory effects of AE on melanoma cell proliferation and invasion power, accompanied by the stimulation of cell differentiation parameters. Cell differentiation correlated with a remarkable increase of the activity of the transamidating form of TG2, with a significative enhancement of cell adhesion and aggregation. Impaired invasion was paralleled by the decrease of the secretion of matrix metalloproteinase-9. SIGNIFICANCE The overall data confirm a remarkable antiproliferative, antimetastatic and differentiative capability of this anthraquinone. Results suggest that AE appears particularly promising for its potential application in the newborn differentiation therapy of cancer.

Targeting tumor cells through chitosan-folate modified microcapsules loaded with camptothecin.

Poly(vinyl alcohol) microcapsules have been tailored as carriers to deliver camptothecin, an anticancer drug poorly soluble in water. The capsules have been reacted with a chitosan--folate complex in order to selectively target cancer cells overexpressing the folic acid receptor. Microcapsules decorated with the chitosan--folate complex have been characterized in their uptake and release of camptothecin, following the absorption band at λ = 370 nm diagnostic of the drug molecule. The selectivity of chitosan-folate microcapsules in targeting cancer cells has been demonstrated by fluorescence microscopy using HeLa cells, overexpressing the folate receptor and NIH3t3 fibroblasts as a negative control. The chitosan--folate microcapsules loaded with camptothecin significantly reduce the proliferation of HeLa tumor cells, while they have a negligible effect on fibroblasts. This work demonstrates that the chitosan--folate microcapsules represent a promising system to selectively target hydrophobic drugs, such as camptothecin, to tumor cells.

Similar antineoplastic effects of nimesulide, a selective COX-2 inhibitor, and prostaglandin E1 on B16-F10 murine melanoma cells.

There is now increasing evidence that a constitutive expression of cyclooxygenase 2 (COX-2) plays a role in the development and progression of malignant ectodermal tumours. In this study, we investigated whether the selective inhibition of COX-2 would be beneficial in melanoma treatment. Nimesulide, a selective inhibitor of COX-2 that causes the breakdown of proinflammatory 2-series prostaglandins (PG2), adversely affected the growth of B16-F10 melanoma cells through the induction of differentiation. The intracellular levels of polyamine, as a proliferation marker, were reduced by the treatment; at the same time, transglutaminase activation and increase in melanin content, as differentiation indicators, were observed. The potential antimetastatic activity of the drug was further shown by means of the Boyden invasion assay and gelatin zymography for metalloproteinase activity. Comparable results were obtained after the treatment of cells with one of the 1-series PGs (PGE1). Therefore, our hypothesis is that the antineoplastic activity observed for nimesulide may be ascribed to intracellular changes in alterations in PG levels.

Protein–polyamine conjugates by transglutaminase 2 as potential markers for antineoplastic screening of natural compounds

The role of post-translational modification of cell proteins with polyamines, a reaction catalyzed by a tissue tranglutaminase (TG, EC 2.3.2.13), in the induction of cell differentiation, represents an intriguing strategy to control cell proliferation and metastatic ability of different tumor cell lines. In this review, we focus our attention on the metabolic aspects of some natural compounds (methylxantines, retinoids and flavonoids) responsible of their antitumor effects exerted through the induction of TG activity in cancer cells.

Transglutaminase-dependent antiproliferative and differentiative properties of nimesulide on B16-F10 mouse melanoma cells.

The aim of this study was to collect evidences on the role of transglutaminase (TG, E.C.2.3.2.13) in the antineoplastic properties exerted by nimesulide (NMS), a non-steroidal anti-inflammatory drug, on murine B16-F10 melanoma cells. Treatment of melanoma cells with nimesulide produces a considerable reduction of cell proliferation, paralleled by a remarkable decrease of the intracellular concentration of polyamines spermidine and spermine. NMS treatment induces cancer cell differentiation, likely through the observed enhancement of TG and tyrosinase activities and increase of melanin production, well known markers of melanocyte differentiation. The overall results highlight the possibility that nimesulide acts as antineoplastic agent likely through the induction of intracellular TG activity.

Transglutaminases: key regulators of cancer metastasis

The ability to metastasize represents the most important characteristic of malignant tumors. The biological details of the metastatic process remain somewhat unknown, due to difficulties in studying tumor cell behaviour with high spatial and temporal resolution in vivo. Several lines of evidence involve transglutaminases (TGs) in the key stages of tumor progression cascade, even though the molecular mechanisms remain controversial. TG expression and activity display a different role in the primary tumor or in metastatic cells. In fact, TG expression is low in the primary tumor mass, but augmented when cells acquire the metastatic phenotype. Nevertheless, in other cases, the use of inducers of TG transamidating activity seems to contrast tumor cell plasticity, migration and invasion. In the following review, the function of TGs in cancer cell migration into the extracellular matrix, adhesion to the capillary endothelium and its basement membrane, invasion and angiogenesis is discussed.

Aloin enhances cisplatin antineoplastic activity in B16-F10 melanoma cells by transglutaminase-induced differentiation

Aloin, a natural anthracycline from aloe plant, is a hydroxyanthraquinone derivative shown to have antitumor properties. This study demonstrated that aloin exerted inhibition of cell proliferation, adhesion and invasion abilities of B16-F10 melanoma cells under non-cytotoxic concentrations. Furthermore, aloin induced melanoma cell differentiation through the enhancement of melanogenesis and transglutaminase activity. To improve the growth-inhibiting effect of anticancer agents, we found that the combined treatment of cells with aloin and low doses of cisplatin increases the antiproliferative activity of aloin. The results suggest that aloin possesses antineoplastic and antimetastatic properties, exerted likely through the induction of melanoma cell differentiation.

Aloe-emodin as antiproliferative and differentiating agent on human U937 monoblastic leukemia cells

AIMS Aloe-emodin (AE), a plant derived anthraquinone, has been shown to have anticancer activity in various human cancer cell lines. We have recently reported that AE possesses a differentiative potential on melanoma cells. The purpose of this study was to investigate the possible modulation of defined markers of monocytic differentiation of AE on human U937 cell line. MAIN METHODS U937 cells differentiation has been confirmed unequivocally by Griess and nitroblue tetrazolium reduction assays, protoporphyrin IX accumulation, expression of CD14 and CD11b surface antigens, phagocytic activity, migration and attachment ability. The effect on polyamine metabolism, apoptosis and cytokine production was also investigated. KEY FINDINGS We showed that AE-treated U937 cells exhibit a noticeably rise in transglutaminase activity. This enhanced enzyme activity correlates with AE-induced growth arrest and differentiation to functionally mature monocytes. SIGNIFICANCE Taken together, the results reported here show that AE can promote the macrophage differentiation of U937 cells, suggesting that this anthraquinone could be a potential candidate as a differentiation-inducing selective agent for therapeutic treatment of leukemia.

Inhibition of cell proliferation, migration and invasion of B16-F10 melanoma cells by α-mangostin

In this study, we have evaluated the potential antineoplastic effects of α-mangostin (α-M), the most representative xanthone in Garcinia mangostana pericarp, on melanoma cell lines. This xanthone markedly inhibits the proliferation of high-metastatic B16-F10 melanoma cells. Furthermore, by deeply analyzing which steps in the metastatic process are influenced by xanthone it was observed that α-M strongly interferes with homotypic aggregation, adhesion, plasticity and invasion ability of B16-F10 cells, probably by the observed reduction of metalloproteinase-9 activity. The antiproliferative and antimetastatic properties of α-M have been established in human SK-MEL-28 and A375 melanoma cells. In order to identify pathways potentially involved in the antineoplastic properties of α-M, a comparative mass spectrometry proteomic approach was employed. These findings may improve our understanding of the molecular mechanisms underlying the anti-cancer effects of α-M on melanoma.

Phytochemicals and protein-polyamine conjugates by transglutaminase as chemopreventive and chemotherapeutic tools in cancer.

Identifying novel chemopreventive and chemotherapeutic agents and targeting them to patients at high risk of developing cancer or following curative treatment may go some way towards improving prognosis. This review examines current knowledge regarding the chemopreventive and chemotherapeutic potential of phytochemicals in cancer. Both in vitro and animal studies demonstrate that several phytochemicals increase the activity of intracellular transglutaminases, a family of enzymes involved in cell differentiation, through the covalent conjugation of polyamine to cellular protein, with promising anti-neoplastic properties. The substantial data available on certain plant secondary metabolites makes a strong case for integrating these safe and well-tolerated agents into clinical practice.