CDK/CCN and CDKI Alterations for Cancer Prognosis and Therapeutic Predictivity

The regulation of cell growth and division occurs in an accurate sequential manner. It is dictated by the accumulation of cyclins (CCNs) and cyclin-dependent kinases (CDKs) complexes and degradation of CCNs. In human tumors, instead, the cell cycle is deregulated, causing absence of differentiation and aberrant cell growth. Oncogenic alterations of CCNs, CDKs, and CDKIs have been reported in more than 90% of human cancers, and the most frequent are those related to the G1 phase. Several molecular mechanisms, including gene overexpression, chromosomal translocations, point mutations, insertions and deletions, missense and frame shift mutation, splicing, or methylation, may be responsible for these alterations. The cell cycle regulators are involved in tumor progression given their association with cancers characterized by higher incidence of relapses and chemotherapy resistance. In the last decade anticancer drug researches focused on new compounds, able to target molecules related to changes in genes associated with tumor status. Recently, the studies have focused on the restoration of cell cycle control modulating molecular targets involved in cancer-cell alterations. This paper aims to correlate alterations of cell cycle regulators with human cancers and therapeutic responsivity

effects of a new human recombinant mnsod in the treatment of photoaging and actinic keratosis

Physiological processes, as aerobic metabolism and inflammatory response, generate reactive oxygen species (ROS) that may induce cellular injury when their amount is increased and antioxidant defense mechanisms are overwhelmed. Also, ROS are generated following UV skin irradiation able to deplete the natural antioxidant defenses in the skin. The increase in exposure to UV may lead to photoaging and precancerous skin lesions (actinic keratosis). New antioxidant strategies in the prevention and therapy of skin lesions are urgently needed. In this study, we evaluated the antioxidant efficacy of a recombinant form of human manganese superoxide dismutase able to inhibit reactive oxygen species production in some patients affected by severe photoaging and actinic keratosis

CDK/CCN and CDKI Alterations for Cancer Prognosis and Therapeutic Predictivity

The regulation of cell growth and division occurs in an accurate sequential manner. It is dictated by the accumulation of cyclins (CCNs) and cyclin-dependent kinases (CDKs) complexes and degradation of CCNs. In human tumors, instead, the cell cycle is deregulated, causing absence of differentiation and aberrant cell growth. Oncogenic alterations of CCNs, CDKs, and CDKIs have been reported in more than 90% of human cancers, and the most frequent are those related to the G1 phase. Several molecular mechanisms, including gene overexpression, chromosomal translocations, point mutations, insertions and deletions, missense and frame shift mutation, splicing, or methylation, may be responsible for these alterations. The cell cycle regulators are involved in tumor progression given their association with cancers characterized by higher incidence of relapses and chemotherapy resistance. In the last decade anticancer drug researches focused on new compounds, able to target molecules related to changes in genes associated with tumor status. Recently, the studies have focused on the restoration of cell cycle control modulating molecular targets involved in cancer-cell alterations. This paper aims to correlate alterations of cell cycle regulators with human cancers and therapeutic responsivity

The Functional Role of MnSOD as a Biomarker of Human Diseases and Therapeutic Potential of a New Isoform of a Human Recombinant MnSOD

Reactive oxygen species (ROS) are generated as a consequence of metabolic reactions in the mitochondria of eukaryotic cells. This work describes the role of the manganese superoxide dismutase (MnSOD) as a biomarker of different human diseases and proposes a new therapeutic application for the prevention of cancer and its treatment. The paper also describes how a new form of human MnSOD was discovered, its initial application, and its clinical potentials. The MnSOD isolated from a human liposarcoma cell line (LSA) was able to kill cancer cells expressing estrogen receptors, but it did not have cytotoxic effects on normal cells. Together with its oncotoxic activity, the recombinant MnSOD (rMnSOD) exerts a radioprotective effect on normal cells irradiated with X-rays. The rMnSOD is characterized by the presence of a leader peptide, which allows the protein to enter cells: this unique property can be used in the radiodiagnosis of cancer or chemotherapy, conjugating radioactive substances or chemotherapic drugs to the leader peptide of the MnSOD. Compared to traditional chemotherapic agents, the drugs conjugated with the leader peptide of MnSOD can selectively reach and enter cancer cells, thus reducing the side effects of traditional treatments

Effects of a New Human Recombinant MnSOD in the Treatment of Photoaging and Actinic Keratosis

Physiological processes, as aerobic metabolism and inflammatory response, generate reactive oxygen species (ROS) that may induce cellular injury when their amount is increased and antioxidant defense mechanisms are overwhelmed. Also, ROS are generated following UV skin irradiation able to deplete the natural antioxidant defenses in the skin. The increase in exposure to UV may lead to photoaging and precancerous skin lesions (actinic keratosis). New antioxidant strategies in the prevention and therapy of skin lesions are urgently needed. In this study, we evaluated the antioxidant efficacy of a recombinant form of human manganese superoxide dismutase able to inhibit reactive oxygen species production in some patients affected by severe photoaging and actinic keratosis

Effects of a New Human Recombinant MnSOD in the Treatment of Photoaging and Actinic Keratosis.

Physiological processes, as aerobic metabolism and inflammatory response, generate reactive oxygen species (ROS) that may induce cellular injury when their amount is increased and antioxidant defense mechanisms are overwhelmed. Also, ROS are generated following UV skin irradiation able to deplete the natural antioxidant defenses in the skin. The increase in exposure to UV may lead to photoaging and precancerous skin lesions (actinic keratosis). New antioxidant strategies in the prevention and therapy of skin lesions are urgently needed. In this study, we evaluated the antioxidant efficacy of a recombinant form of human manganese superoxide dismutase able to inhibit reactive oxygen species pro- duction in some patients affected by severe photoaging and actinic keratosi

The leader peptide of a human rec. MnSOD as molecular carrier which delivers high amounts of cisplatin into tumor cells inducing a fast apoptosis in vitro

The leader peptide of a recombinant MnSOD (rMnSOD-Lp) constitutes the carrier that allows rMnSOD to penetrate tumor cells. A synthetic preparation of rMnSOD-Lp was \u2076\u2078Ga labeled (rMnSOD-Lp- \u2076\u2078Ga) and injected into animals bearing spontaneous mammary cancers, followed by PET examinations, which demonstrated unambiguously the tumor sites in all the animals, suggesting that if rMnSOD-Lp was able to transport the radioisotope into tumor cells, it would also be able to deliver cytotoxic molecules. The rMnSOD-Lp was, therefore, conjugated to cisplatin (rMnSOD-Lp-CC) and added to cultured tumor cells. Equal concentrations of cisplatin were used for the tests. After treating the ovarian cancer cells with 11.1 \u3bcg of cisplatin alone, analysis by atomic absorbance spectrophotometry was able to detect only 6 ng of platinum, whereas when the same cells were treated with the same amount of cisplatin conjugated to leader peptide rMnSOD, 387 ng of platinum were detected, i.e., an amount 80 times greater. Only the tumor cells died following treatment with rMnSOD-Lp-CC; molecular analysis revealed that its addition generated an increasing expression of Erk-2 and Bax products, which could be inhibited only by a selective MAP/ERK kinase inhibitor (PD98059), revealing that rMnSOD-Lp-CC has an apoptotic function, exactly as occurs when using the cisplatin alone. Data are statistically significant and indicate that by using rMnSOD-Lp-CC, the cisplatin can be transformed from an agent with antireplicative activity into a specific and selective antitumor molecule, increasing its therapeutic index. We think that rMnSOD-Lp-CC deserves to be considered as a new antitumor agent

Effetti della rMnSOD nella terapia delle discheratosi attiniche

Gli UV sono in grado di indurre modificazioni sia quantitative che qualitative delle cellule immunocompetenti e possono alterare l’immuno-sorveglianza verso i tumori cutanei. L’irradiazione ultravioletta stimola i cheratinociti a secernere citochine e fattori di crescita che intervengono sulle cellule deputate alla risposta immunitaria (Santoianni, 2003 ). Dati sperimentali, epidemiologici e clinici negli ultimi anni hanno messo in evidenza l’importanza dell’ultravioletto lungo nella genesi di cheratosi attiniche (Nino 2006). Organismi esposti a radiazioni ionizzanti sono principalmente danneggiati dai radicali liberi, generati dalla radiolisi dell'acqua contenuta nelle cellule. Inoltre è stata dimostrata una significativa riduzione del danno tissutale da irradiazione utilizzando trattamenti con una manganese superossido dismutasi. Le superossido dismutasi (SOD) sono enzimi che hanno un ruolo chiave nella prevenzione di tutte le patologie determinate dal danno ossidativo (Borrelli et al., 2009). Le SOD sono implicate nella difesa antiossidante di quasi tutte le cellule, poiché catalizzano la dismutazione del radicale superossido a perossido d’idrogeno che, successivamente, è convertito in ossigeno e acqua dall’enzima catalasi. Questi enzimi hanno la capacità di prevenire il danno provocato dagli alti livelli di ROS prodotti, in particolare da radiazioni ionizzanti (Epperly et al., 2003). Nelle cellule sono presenti tre isoforme: la SOD 1 è localizzata nel citosol, la SOD 2 nella matrice mitocondriale, mentre la SOD 3 è secreta nello spazio extracellulare (Oberley, 2005). L’isoforma SOD 2, conosciuta anche come manganese superossido dismutasi (MnSOD), ricopre un ruolo di notevole importanza nella conversione del radicale superossido (O2.-) nei mitocondri e, dunque, rappresenta la prima linea di difesa contro questo radicale (Wang et al., 2001). Recentemente nel laboratorio di ricerca del dott. Aldo Mancini (Istituto Nazionale Tumori G.Pascale) è stata isolata un’isoforma di SOD da cellule di liposarcoma umano in coltura (LSA-Type MnSOD) che ha mostrato sia in vivo che in vitro una azione citotossica specifica e selettiva solo per le cellule esprimenti il recettore per gli estrogeni. Pur avendo la stessa attività enzimatica comune a tutte le SOD, la LSA-Type MnSOD si differenzia dalla sua corrispondente nativa per la presenza del peptide leader (da 24 amminoacidi), evidentemente non clivato, che le conferisce la peculiare capacità di penetrare in tutte le cellule. Tale proteina è stata riprodotta in forma ricombinante, rMnSOD, a partire da uno specifico clone di cDNA derivato da cellule di liposarcoma umano (Mancini et al., 2006). La rMnSOD è risultata, in vitro, essere radioprotettiva per le cellule normali e radiosensibilizzante per quelle tumorali. Inoltre, animali sani, esposti a dosi letali di radiazioni ionizzanti in presenza della rMnSOD (1,4 µM), mediante iniezioni s.c. sono sopravvissuti al danno radiante rimanendo vivi 30 giorni dopo l'irradiazione, tempo in cui è stato interrotta il controllo della loro sopravvivenza. Al contrario, animali irradiati con le stessse dosi letali, in assenza della rMnSOD, morivano dopo 7-8 giorni dall’irradiazione (Borrelli et al.2009). La notevole capacità enzimatica della rMnSOD, in formulazione topica, di neutralizzare i radicali liberi che incontra e che si accumulano nei tessuti danneggiati da qualsivoglia noxa patogena, è stata dimostrata, inoltre, con il suo utilizzo nella cura di una necrosi profonda ed estesa a testa, collo, e natatoie di un esemplare di tartaruga marina Caretta caretta. Il trattamento topico di tali esemplari con rMnSOD ha consentito una piena restitutio ad integrum anche nei siti delle necrosi che avevano provocato esposizione dell'osso (Occhiello A. et al 2009). Sulla base dei risultati precedentemente ottenuti è stato allestito uno studio su pazienti afferenti alla Dermatologia dell'Ospedale Ascalesi di Napoli, i quali si sono spontaneamente dichiarati disponibili al trattamento con la formulazione cosmetica contenente la rMnSOD, Sono stati arruolati per lo studio 30 pazienti di entrambi i sessi di eta’ compresa tra i 35 e i 70 anni con fotodanno medio severo con presenza di discheratosi attiniche. I pazienti hanno praticato terapia topica con una formulazione O/A a base di rMnSOD, mattina e sera per due mesi. Ogni settimana è stato effettuato un controllo e sono state fotografate le lesioni per monitorare l'effetto della terapia. Già nella prima settimana è stato osservata una diminuzione dello stato infiammatorio in tutti i pazienti trattati. A due mesi di trattamento è stato osservato miglioramento consistente del fotodanno, migliorata la compattezza e la luminosità della cute con riduzione della elastosi solare e scomparsa o regressione parziale delle discheratosi. La formulazione topica della rMnSOD merita di essere considerata come un promettente farmaco con potente azione antiinfiammatoria utilizzabile in dermatologia. • Borrelli A, Schiattarella A, Mancini R, Morrica B, Cerciello V, Mormile M, d'Alesio V, Bottalico L, Morelli F, D'Armiento M, D'Armiento FP, Mancini A. Free Radical Biology and Medicine. 2009 Jan 1;46(1):110-6. Epub 2008 . • Epperly M.W., Gretton J. E., Sikora C.A., Jefferson M., Bernarding M., Nie S. and Greenberger J.S. (2003). Radiation Research. 160 (5): 568-578. • Mancini A., Borrelli A., Schiattarella A., Fasano S., Occhiello A., Pica A., Sher P., Tommasino M., Nüesch J. P. F. and Rommelaere J. (2006). International Journal of Cancer. 119 (4): 932-943. • Oberley L.W. Biomedicine & Pharmacotherapy. 59 (4): 143-148. • Occhiello A., Bentivegna F., Borrelli A., Schiattarella A., Mancini A., Pica A. Comparative clinical Pathology. DOI 10.1007/s00580-009-0816-9 • Wang L.I., Miller D.P., Sai Y., Liu G., Su L., Wain J.C., Lynch T.J. and Christiani D.C. (2001). Journal of the National Cancer Institute. 93 (23): 1818-1821. • Santoianni P., Nino M. Giornale italiano di dermatologia e venereologia. 2003. 138(6):455-64 • Nino M., Santoianni P. . Giornale italiano di dermatologia e venereologia 2006. 141(5):471-

A new perspective for the treatment of human endometrial cancer

The endometrial cancer is one of the principal causes of death in women (Siegel et al., 2012). The main treatment for this cancer is hysterectomy that involves loss of fertility (Holland, 2008). If the tumor is not well defined and metastasizes, radio/chemo therapy are needed. The survival after chemotherapy is about 5-10 years, because it cannot kill all tumor cells. So, an isoform of manganese superoxide dismutase isolated from a human liposarcoma cell line, obtained as recombinant molecule (rMnSOD), which displayed oncotoxic action on cultured breast cancer cells (Mancini et al.,2006), was used to treat human endometrial cancer cells. This enzyme catalizes the dismutation of O2 – free radicals in H2O2, preventing the accumulation of ROS. H2O2 can be further converted into H2O and O2 by catalase and ⁄or glutathione peroxidase. Once rMnSOD penetrates cancer cells, it transforms free radicals into H2O2. This, in neoplastic cells, may lead to an high accumulation of H2O2 that causes apoptosis of them, because of their lower level of catalase (Mancini et al., 2008). We evaluated the effects of the rMnSOD treatment on cultured human endometrial adenocarcinoma cell line HTB-112. We demonstrated the oncotoxic effect of this protein, which lead HTB/112 cells to apoptotic death, by immunocytochemistry at light and electron microscopy and by comet assay test, which shows early DNA damage induced by the accumulation of H2O2. References: Siegel CA 62, 10–29; 2012; Holland Clin Oncol 20:448-56 2008 ; Mancini Int J Cancer, 119: 932–943¸ 2006