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In the present study, we investigated the effect of apigenin, a flavonoid widely present in fruits and vegetables, on a tongue oral cancer-derived cell line (SCC-25) and on a keratinocyte cell line (HaCaT), with the aim of unveiling its antiproliferative mechanisms. The effect of apigenin on cell growth was evaluated by MTT assay, while apoptosis was investigated by phosphatidyl serine membrane translocation and cell cycle distribution by propidium iodide DNA staining through flow cytometry. In addition the expression of cyclins and cyclin-dependent kinases was evaluated by western blotting. A reduction of apigenin-induced cell growth was found in both cell lines, although SCC-25 cells were significantly more sensitive than the immortalized keratinocytes, HaCaT. Moreover, apigenin induced apoptosis and modulated the cell cycle in SCC-25 cells. Apigenin treatment resulted in cell cycle arrest at both G0/G1 and G2/M checkpoints, while western blot analysis revealed the decreased expression of cyclin D1 and E, and inactivation of CDK1 upon apigenin treatment. These results demonstrate the anticancer potential of apigenin in an oral squamous cell carcinoma cell line, suggesting that it may be a very promising chemopreventive agent due to its cancer cell cytotoxic activity and its ability to act as a cell cycle modulating agent at multiple levels.

Epidemiological data consistently demonstrate a reduced cancer risk associated with a polyphenols rich diet. Hibiscus sabdarifa (HS), a polyphenols rich plant widely consumed worldwide as beverage and used in folk medicine, has recently gained interest thanks to its antioxidant, anti-inflammatory and chemopreventive properties. In the present study we investigated the antitumoral potential of HS extract in two different human tumor cell lines: Multiple Myeloma cells (RPMI 8226) and Oral Squamous Cell Carcinoma cells (SCC-25). MTT assays showed that HS extract induced a dose-dependent viability re-duction in both the cells lines. For the subsequent experiments we used HS at the concentration of 5 mg/ml that was the most effective in inducing cell viability reduction after 48h of treatment. Viable cell count using trypan blue staining demonstrated that the HS extract induced decrease in cell growth of both the cell lines and this was due to a reversible cytostatic rather than a cytotoxic effect. Wound-healing and cell invasion assays, respectively performed by a scratch of cell monolayer and Boyden Chamber transwell test, demonstrated that HS ex-tract was able to reduce motility and invasiveness in both RPMI 8226 and SCC-25 cells. The chemical inhibition of ERK1/ERK2 and PI3K, with U0126 and wortmannin re-spectively, reduces proliferation and migration of both SSC-25 and RPMI cells and HB extract treatment played an additive action with the inhibitors. In conclusion, our results suggest that HS extract have antitumoral properties, since it proved to inhibit tumoral cell growth and cell migration and invasiveness. It is interesting to note that HS extract is effective against two very different tumor cell lines. In fact, RPMI 8226 cells are of hematopoietic origin and grow in suspension, whereas SCC-25 cells derive from epithelium and are characterized by adherent cell growth. Therefore, although further studies are needed to clarify the molecular mechanisms involved i

Oral Cancer (OC) is one of the most frequent cancer in Head and Neck district and Oral Squamous Cell Carcinoma (OSCC) constitutes the large majority of the neoplasia arising in oral cavity. OSCC remains a hampering matters for clinics, since the overall disease free survival has not significantly increased during the last decades and invasion to surrounding tissue and to regional lymph nodes is often reported. Therefore new strategies to prevent and inhibit OSCC growth and invasion are highly desirable and new therapeutic approaches are currently tempted also with the use of natural compounds. Myricetin (MYR) and Naringenin (NAR), two naturally occurring flavonoids, widely diffused in plants, fruits and vegetable, have recently gained consideration thanks to their anti oxidant, anti inflammatory and anti tumoral properties. In this study their potential anticancer effect has been evaluated on an OSCC cell line, SCC-25 and on spontaneously immortalized non tumoral keratinocytes, HaCaT cells. MYR and NAR induce a significant cell growth inhibition in SCC-25 cells, in addition NAR selectively affected cancer cells, since it does not impair HaCaT cell growth. Furthermore an additive effect of MYR and NAR has been highlighted. The cell proliferation inhibition is not related to apoptosis induction, as demonstrated by evaluation of phosphatidyl serine membrane translocation and dapi staining. On the contrary MYR and NAR effect depends on the cell cycle progression impairment. Wound-healing and cell invasion assays, respectively performed by cell monolayer scratch and Boyden Chamber transwell test, demonstrate that the two flavonoids are able to reduce motility and invasiveness on both SCC-25 and HaCaT cells. In conclusion the results of the present study show the anticancer potential of NAR and MYR on OSCC, since both flavonoids prevent cancer cell proliferation through a cytostatic effect, by the impairment of cell cycle progression. Moreover both the flavonoids inhibit

Cisplatin is an antineoplastic drug widely used for the treatment of several solid tumours. However, the side effects related to cisplatin-based anticancer therapy often outweigh the benefits. Therefore, the identification of new anticancer strategies able to offer a better toxicity profile while maintaining the same level of efficacy as platinum-based treatments would be highly desirable. We assessed the efficacy of synchrotron radiation in triggering the Auger effect in human A549 non-small cell lung cancer and IGROV-1 ovarian cancer cells pre-treated with cisplatin. Cisplatin was chosen as the carrier of platinum atoms in the cells because of its alkylating-like activity and the irradiation was done with monochromatic beams above and below the platinum K-shell edge (78.39 keV). On cisplatin-treated cells, at concentrations allowing 80 percent of cell survival with respect to controls, no differences were observed in cell viability when they were irradiated either above or below the K-shell edge of platinum, suggesting that cisplatin toxicity can mask the enhancement of cell death induced by the irradiation. At lower cisplatin concentrations allowing 95-90 percent of cell survival, an enhancement in cellular death with respect to conventional irradiation conditions was clearly observed in all cancer types when cells were irradiated with beams either above or below the platinum K-shell edge. Our results lend additional support to the suggestion that the Photon Activation Therapy in combination with cisplatin treatment should be further explored in relevant in vivo models of glioma and non-glioma cancer models

Neurotoxicity is a common dose-limiting side-effect of several drugs (Cavaletti et al., 2008). So far a validated test method to screen drugs neurotoxicity does not exist, therefore in this interdepartment study we have analyzed the effectiveness of a morphometric neurotoxicty assessment model. Drug neurotoxicity evaluation is based on embryonic rat dorsal root ganglia (DRG) organotypic culture. DRG primary sensory neurons are the principal target of drugs neurotoxic action. In fact, primary sensory neurons lie outside the blood-nerve barrier and are supplied by capillaries with fenestrated walls. Moreover, the axons of these cells are among the longest of the entire nervous system and, therefore, are more susceptible to any agent that interferes with the energy metabolism or the structural basis of axonal transport. In particular, in this interdepartment study, the interference of the under study neurotoxic compound with NGF-induced neurite elongation is analysed. The effectiveness and reproducibility of this model, even if commonly used to test drugs, has not yet been demonstrated. In order to assess the validity of this in vitro model, antineoplastic drugs known to be in clinical use and in animal models neurotoxic (paclitaxel and oxaliplatin) or not dangerous (cyclophosphamide and 5-Fluorouracil) have been tested. DRGs explanted from E15 rat embryos have been treated for 24h with drugs concentrations comparable to those achievable in vivo. The length of the longest neurite of each DRG has been measured by ImageJ program. Experiments have been performed by three different blinded researchers in two different laboratories. Mean and standard deviation of each experiment were obtained, subsequently the mean value and standard deviation of the three independent experiments for each researcher were calculated. Data obtained by the three researchers in two different laboratories resulted statistically comparable and no significant differences were detected (One Way Ano

Cisplatin is an antineoplastic drug widely used in clinic for the treatment of several solid tumours. However, at times, the side effects related to cisplatin-based anticancer therapy often outweigh the benefits. Several of these side effects are now manageable with effective treatment planning and supportive care strategies (e.g. anti-emetics, hematopoietics growth factors, physical treatments, hyperhydration), but others are still unsolved, can be dose-limiting and represent an unmet clinical need, for instance nervous system damage. Therefore, the identification of new anticancer strategies able to offer a better toxicity profile maintaining the same level of efficacy as platinum-based treatments would be highly desirable. We tested the efficacy of synchrotron radiation to trigger the Auger effect in the living human A549 non-small cell lung cancer (NSCLC) and IGROV-1 ovarian cancer cells pre-treated with cisplatin. Furthermore we tested the anticancer treatment in human VIPI glioblastoma stem cells that seem to be one of the most important determinants in the extraordinary resistance of GBL to any treatment tested so far. The experiments were carried out at the ID17 beamline of the European Synchrotron Radiation Facility, where a high-fluence monochromatic beam adjustable from 20 to 80 keV is available. Cisplatin was chosen as the carrier of platinum atoms in the cells because of its alkylating-like activity and the irradiation was done with monochromatic beams above and below the platinum K-shell edge (78.39 keV). Control cell survival curves were comparable with those obtained for the same cells under conventional irradiation conditions. On cisplatin-treated cells at a concentrations inducing 80% of cell survival with respect to the control, no differences were observed in cell survival when cells were irradiated either above or below the K-shell edge of platinum, suggesting that cisplatin toxicity can mask the enhancement of cell death induced by the irradiat

Cisplatin is the archetypal inorganic drug and it is nowadays one of the most efficient metal-based anticancer agents, targeting a large number of solid tumours. Although highly effective in treating a variety of cancers, cisplatin treatment is still limited by severe side effects such as neuro-, hepato- and nephro-toxicity and by inherited or acquired resistance phenomena, only partially overcome by the use of new platinum drugs (i.e. oxaliplatin and carboplatin). These problems have stimulated the research and development of alternative therapeutic strategies based on different heavy metals. In this work we investigated the neurotoxicity effect of two new promising water soluble anticancer complexes based on copper [Cu(PTA)4PF6] and gold [Au(PTA)4PF6]. Cisplatin and oxaliplatin neurotoxicity was also measured as reference drugs. Neurotoxicity was evaluated by an in vitro model based on the ability of NGF exposed embryonic rat dorsal root ganglia (DRG) organotypic culture to grow neurite. In particular the interference of the under study neurotoxic compound with neurite elongation was analysed. DRGs explanted from E15 rat embryos were treated for 48h with different drug concentrations including the IC50 evaluated for different tumour cell lines. In our model cisplatin and oxaliplatin resulted neurotoxic (inducing a reduction of neurite outgrowth more than 50%) at concentrations achievable in plasma of patients treated with the same drugs confirming data obtained in clinical studies. On the contrary, the neurotoxicity assay demonstrated that copper-based compound was neurotoxic at higher concentrations with respect to the IC50 obtained for all tumour cell lines tested. On the other hand preliminary neurotoxicity data suggest that gold-based compound is neurotoxic at lower concentration than IC50 calculated for almost tested cancer cell lines. Our results, together with the low IC50 of the copper compound compared to the one observed for cisplatin and oxaliplatin, su

The peripheral nervous system is a frequent target of toxic agents. The accurate identification of the sites of neurotoxic action through the morphological characterization of reliable in vivo models or in vitro systems can give fundamental clues when investigating the pathogenesis and interpreting the clinical features of drug-induced neuropathy. The morphological approach has been used to investigate almost all the anticancer drugs able to induce chemotherapy-induced peripheral neurotoxicity, i.e. platinum drugs, antitubulins and proteasome inhibitors. No models have ever been described for thalidomide. This review demonstrates that any pathogenetic study on chemotherapy-induced peripheral neurotoxicity must be based on solid morphological observations obtained in reliable animal and in vitro models. This is particularly true in this setting, since the availability of tissues of human origin is extremely limited. In fact, peripheral (generally sural) nerve biopsies are never required for diagnostic purposes in chemotherapy-treated cancer patients, and their use for a purely scientific aim, although potentially very informative, is not ethical. Moreover, several neurotoxic drugs target the dorsal root ganglia neurons, and it is very difficult to obtain high-quality specimens even from early autopsies. It is, therefore, our opinion that an extensive morphological assessment of the in vitro and in vivo effect of any potentially neurotoxic antineoplastic drugs, as well as of neuroprotectant agents, should be taken into consideration right from the earliest stages of their development

Nowadays obesity and its related diseases represent a major health problem with an increasing worldwide prevalence. Hyperplasia and hypertrophy of adipocytes lead to an excessive fat accumulation that is not efficiently prevented by current pharmacological treatments. So the research on anti-obesity drugs with good efficacy and tolerability able both to prevent and to reduce fat accumulation is of pivotal interest. In the present study we evaluated in vitro the effects of Valproic Acid, Berberin and Resveratrol on adipogenesis. Our experimental model was represented by human Mesenchymal Stem Cells (hMSCs), physiological precursors of adipocytes that can differentiate into adipocytes also in vitro. Preliminary cytotoxicity assays were performed in order to choose non-toxic doses of the three drugs. hMSCs were induced to adipogenic differentiation and treated with Valproic Acid, Berberin and Resveratrol at the selected doses. Controls were represented by hMSCs treated for adipogenesis in absence of the drugs. At different time points intracellular lipid droplets accumulation, a typical feature of adipogenesis, was assessed by Oil Red O staining. Valproic Acid, Berberin and Resveratrol inhibited hMSCs adipogenic differentiation in a dose dependent manner as demonstrated by the reduction of the lipid droplets accumulation. To understand the molecular mechanisms of the drugs-induced adipogenesis inhibition, we focused our attention on the effects of the drugs treatment on cell cycle progression, known to be altered by many antiadipogenic drugs, and on the MAP Kinases ERK1 and ERK2, involved in the adipogenesis control. We evaluated the expression of cyclins and CDKs by immunoblotting and flow-cytometry analyses, demonstrating that Valproic Acid, Berberin and Resveratrol interfere on cell cycle progression. The expression and the phosphorylation status of the two kinases ERK1 and ERK2 were assessed by immunoblotting demonstrating an increase of ERK1 phosphorylation (i.e.

Neurotoxicity is a common dose-limiting side-effect of several drugs. So far a validated test method to screen drugs neurotoxicity does not exist, therefore in this interdepartment study we have analyzed the effectiveness of a neurotoxicty assessment model. Drug neurotoxicity evaluation in this model is based on the ability of NGF exposed embryonic rat dorsal root ganglia (DRG) organotypic culture to grow neurite; in particular the interference of the under study neurotoxic compound with neurite elongation is analysed. The effectiveness and reproducibility of this model , even if commonly used to test drugs , has not yet been demonstrated. In order to assess the validity of this in vitro model, antineoplastic drugs known to be in clinical use and in animal models neurotoxic (paclitaxel and oxaliplatin) or not dangerous (cyclophosphamide and 5-Fluorouracil) have been tested. DRGs explanted from E15 rat embryos have been treated for 24h with drugs concentrations comparable to those achievable in vivo. The length of the longest neurite of each DRG has been measured by ImageJ program. Experiments have been performed by three different blinded researchers in two different laboratories. Mean and standard deviation of each experiment were obtained, subsequently the mean value and standard deviation of the three independent experiments for each researcher were calculated. Data obtained by the three researchers in two different laboratories resulted statistically comparable and no significant differences were detected (One Way Anova analysis of variance and Tukey post test; p<0.05). This interdepartment in vitro study, therefore, indicates that the DRG organotypic culture represents a reliable model to study drug neurotoxicity. The test method permits to make prediction of neurotoxic effects on humans because the concentrations tested are the same to which DRG are exposed during clinical use. This model will allow to reduce animal testing for screening of new drugs and to de

Organic cation transporters (OCTs) are critical in drug absorption, targeting, and disposition. It has become increasingly clear the role of these transporters in key tissues responsible for drug absorption and disposition: kidney, liver and intestine. However, only limited information is available regarding their distribution and activity in nervous system. Cisplatin (CDDP), a widely used chemotherapeutic agent for the treatment of several solid tumors induces a severe and dose-limiting sensory neuropathy, due to damage of the primary sensory neurons of the dorsal root ganglia (DRG) where it forms DNA adducts. This study was aimed to investigate (1) the expression of different OCTs (OCT-1, OCT-2, OCTN-1 and OCTN-2) in DRG neurons and (2) their putative role in CDDP-induced peripheral neurotoxicity. OCTs expression in rat DRG was studied by TaqMan Real Time PCR in control and CDDP treated (2 mg/Kg i.p. 2qw x4) Wistar rats. It was demonstrated that CDDP administration reduced OCT2 mRNA levels with respect to the control. To examine the functional role of OCT-2, the correlation existing between the neurophysiologic/morphometric parameters and the OCT-2 mRNA levels were studied. This analysis evidenced that the reduction of OCT-2 expression correlated with the decrease of NCV and with the reduction of soma, nucleus and nucleolus size in treated animals. To obtain mechanistic information about the role of OCT-2 in CDDP neurotoxicity a pilot in vivo study was performed. Female Wistar rats were randomized in 3 different groups and treated as follows: CDDP 2 mg/Kg i.p. 2qw x4; CDDP 2 mg/Kg i.p. 2qw x4 + cimetidine (CMT) 60 mg/kg os daily for 4 weeks; un-treated control rats. The OCT-2 inhibitor CMT could not revert the CDDP-induced NVC impairment, although it significantly ameliorated morphometric parameters. An in vitro model of DRG neurons obtained from embryonic rats E15 was used.to confirm a partial neuroprotective effect of CMT. Nerve growth factor (NGF) differentiate

Collagen is ubiquitous in the human body and it is the main component of connective tissue. Collagen is also the principal component of endopelvic fascia and it is involved in the physiopathology of female organ prolapse (POP). The role of collagen in the physiopathology of POP has been studied with conflicting results, being difficult to distinguish the cause from the effect and to standardize the samples. The turnover of connective tissue in the vaginal wall is an important process both in healthy women and in women with prolapse. MMP-1, MMP-8 and MMP-13 specifically cleave native triple collagen (I, II, III) helix, yelding two fragments. These fragments are subject to gelatinase (MMP-2 and MMP-9) degradation. MMP-2 and MMP-9 activity, may be regulated at different levels (trascriptional and activation level) influencing the amount of collagen. Aim of this study is to find a correlation between staging of anterior vaginal wall prolapse and morphologic and biochemical (MMP-2 and MMP-9 amount) features of anterior vaginal wall. Anterior vaginal wall full thickness biopsies (eightyfive patients) including endopelvic fascia have been collected during surgical procedures. Recurrence or vault prolapse have been ruled out. POP samples have been classified according to POP-Quantification and they have been divided in three groups: group A (controls), group B (I and II stage prolapse) and group C (III and IV stage prolapse). Each sample has been processed for both microscopic and zymography analysis. Formalin fixed samples have been embedded, cut and stained with Trichromic Masson Goldner (TMG) stains. Microscopic analysis have been performed to evaluate collagen amount, collagen organization, and to assess muscular component. All parameters have been scored semi-quantitatively by two blind observers on vaginal mucosa and underlying fascia. MMP-2 and MMP-9 activity have been evaluated by gelatine zymography. Statistical evaluation have been made by One-Way ANOVA analysis o

Organic cation transporters (OCTs) play an important role in transporting cationic xeno- and endobiotics across biological membranes. In particular their role in platinum drugs transport, such as cisplatin and oxaliplatin, has recently been studied in many normal tissues and cancer cells. It has become increasingly clear the role of these transporters in key tissues responsible for drug absorption and disposition: kidney, liver and intestine. However, only limited information is available regarding their distribution and activity in nervous system. This study was aimed to investigate (1) the expression of OCT2 in rat DRG and (2) to quantify the developmental changes in the mRNA expression of OCT2 in DRG neurons isolated from embryonic E15 and young adult Wistar rats. OCT2 mRNA expression in rat DRG was localized by in situ hybridization experiments and quantified by TaqMan Real Time PCR. Our results demonstrated the expression of OCT2 mRNA in rat DRG and its neuronal localization. In order to verify possible developmental changes in the OCT2 expression we quantified by TaqMan Real Time PCR the OCT2 mRNA using in vitro models of DRG neurons obtained from embryonic E15 rats and from young adult rats. OCT2 mRNA expression was lower in embryonic neuron cultures with respect to the one obtained from young adult rats, demonstrating a developmental change in the expression of this transporter in rat DRG neurons. The OCT2 mRNA expression in 4 weeks of age rats approached adult expression levels. Furthermore in literature is reported that the OCT2 mRNA expression in the kidney is gender dependent. Our preliminary data obtained in DRG of adult Wistar rats did not demonstrate the gender difference in OCT2 mRNA expression observed in the kidney. Key words: organic cation transporter, dorsal root ganglia neurons, developmental expression Supported in part by an unrestricted research grant from the “Fondazione Banca del Monte di Lombardia”

Advances in clinical treatment of head and neck squamous cell carcinoma (HNSCC) are hampered by its high infiltrative potential leading to distal metastasis. Since their ability to degrade the basal lamina and extracellular matrix, matrix metalloproteinases (MMP) have a pivotal role in tumor invasion. The overexpression and the aberrant activity of MMPs especially of MMP2 and MMP9, during HNSCC development and progression have been reported. However, up to now little is known about the mechanism of their regulation in HNSCC. It has been demonstrated that MMP2/9 expression is negative regulated by extracellular signal regulated kinase 1 and 2 (ERK1/2) in HNSCC cell lines. ERKs are protein kinases belonging to the mitogen-activated protein kinases family, and they are involved in the regulation of different cellular aspects, from apoptosis to cell proliferation and differentiation. In the present study we evaluated MMP2 and MMP9 activity by gelatine zymography in 16 tissue samples of HNSCC and their paired normal mucosa from patients undergoing surgical treatment. Moreover, ERK1/2 activation was analyzed by immunoblotting. A statistically significant decrease in the levels of activated ERK2 in cancer specimens in comparison with paired normal tissues was observed, whereas a significant increase in the activity of MMP2 was found in cancer specimens. However, the statistical analysis failed to demonstrate a correlation between the increase in MMP2 activity and the reduction of ERK1/2 activation levels. The results obtained, therefore, rule out, for the first time in an ex vivo study, the existence of a negative correlation between ERK1/2 activation and MMP2 activity.

Chemotherapy-induced peripheral neurotoxicity (CIPN) is a side effect limiting cisplatin (CDDP) and docetaxel (DOCE) treatment. Erythropoietin (EPO) is a hematopoietic growth factor also displaying neurotrophic properties. Evidence suggests that EPO's neuroprotective action may rely on PI3K/AKT pathway activation; however, data regarding the EPO neuroprotective mechanism are still limited. This study evaluated the effect of EPO on organotypic cultures of rat dorsal root ganglia (DRG) and in primary cultures of DRG-dissociated sensory neurons exposed to CDDP- and DOCE-induced neurotoxicity, aiming to investigate EPO's neuroprotective mechanism. Subsequently, the levels of AKT expression and activation were analyzed by Western blot in neurons exposed to CDDP or DOCE; AKT's role was further evaluated by using a chemical inhibitor of AKT activation, wortmannin. In these models EPO, was protective against both CDDP- and DOCE-induced cell death and against CDDP-induced neurite elongation reduction. A modulation of AKT activation was observed in CDDP-treated neurons, and the presence of wortmannin prevented EPO's neuroprotective action against CDDP toxicity but did not have any effect on EPO's protection against DOCE-induced toxicity, thus ruling out the PI3K-AKT pathway as the mechanism of EPO's effect in neuronal death prevention after DOCE exposure. Our results confirm in vitro the effectiveness of EPO as a neuroprotectant against both CDDP- and DOCE-induced neurotoxicity. In addition, a role of PI3K/AKT in EPO's protection against CDDP, but not against DOCE, neurotoxicity was shown, suggesting that alternative pathways could be involved in EPO's neuroprotective activity. (

Epothilones are a novel class of microtubule-targeting anticancer agents that are neurotoxic. In this study, we investigated the epothilone B toxic effect in vitro and we characterized in vivo the general and neurological side effects of epothilone B administration in Wistar and Fischer rats. The in vitro experiments made it possible to explore a wide concentration range (0.1 nM-1 muM) and evidenced a dose-dependent effect of epothilone B exposure on neuron neurite elongation. This dose-dependent neurotoxic effect was confirmed in both in vivo studies performed on two different rat strains at the neurophysiological, behavioral and pathological levels in the dose range 0.25-1.5 mg/kg iv weekly x 4 weeks and tubulin hyper-polymerization was demonstrated in sciatic nerve specimens. These are the first studies of the neurological effects of epothilone B and they can provide a basis for extending pre-clinical investigation to other members of the epothilone family

Organic cation transporters (OCTs) are critical in drug absorption, targeting, and disposition. It has become increasingly clear that multiple mechanisms are involved in organic cation transport in the key tissues responsible for drug absorption and disposition: the kidney, liver and intestine. However, only limited information is available regarding their distribution and activity in the central and peripheral nervous system. Cisplatin (CDDP), a widely used chemotherapeutic agent for the treatment of several solid tumors induces a severe and dose-limiting sensory neuropathy, due to damage of the primary sensory neurons of the dorsal root ganglia (DRG) where it forms DNA adducts. Previous in vivo and in vitro studies demonstrated that OCTs are implicated in CDDP nephrotoxicity. The present study was aimed to investigate (1) the presence of different OCTs (OCT-1, OCT-2, OCTN-1 and OCTN-2) in embryonic and adult rat DRG neurons and (2) their putative role in CDDP induced neurotoxicity. To examine the functional role of OCT-2, an in vivo study investigated the correlation existing between the neurophysiologic/morphometric parameters and the OCT-2 mRNA levels when rats were treated with CDDP. This analysis evidenced that the reduction of OCT-2 expression correlated with the decrease of NCV as well as with the reduction of soma, nucleus and nucleolus size in treated animals. Finally, to obtain mechanistic information about the role of OCT-2 in CDDP neurotoxicity we used an in vitro model of DRG neurons obtained from embryonic rats E15. The OCTs expression in rat DRG was studied by TaqMan Real Time PCR and it was demonstrated that CDDP administration reduced OCT-2 mRNA levels with respect to the control. In order to verify the involvement of OCT2 in the neuronal uptake of CDDP, nerve growth factor (NGF) differentiated neurons were exposed to CDDP alone or in combination with the OCT-2 inhibitor cimetidine (CMT). After 24 and 48 hours of co-treatment with CDDP and CMT 100