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Nucleostemin gene silencing by siRNA induces differentiation and growth inhibition of NB4 leukemia cell line
S182
Abstracts
a
Department of Clinical Biochemistry, Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Hezarjarib Ave, Isfahan, Iran b Applied Physiology Research Center, Department of Physiology, Isfahan University of Medical Sciences, Isfahan, Iran E-mail addresses: [email protected] (N.M. Masoud), [email protected] (M. Ahmad), [email protected] (H.J. Shaghayegh)
Conclusion: Based on differentiating and growth inhibitory effects of NS-siRNA, NS can be proposed as a new target for treatment of APL patients.
Introduction: Oxidative stress is a causative factor of endothelial dysfunction and plays an important role in the pathophysiology of several vascular diseases. L-serine (L-ser) is known to exert cytoprotection in hepatocytes against oxidative stress. The present study investigates the effect of L-ser on H2O2-mediated cytotoxicity in cultured human umbilical vien endothelial cells (HUVEC). Materials and methods: In this study HUVEC were treated with different concentrations of L-Ser (0.1-3.2 mM). Then, Heme Oxygenase1(HO-1) and Nuclear factor-erythroid 2-related factor 2(Nrf2) levels were evaluated through Sandwich ELISA and DNA-binding based techniques respectively. Furthermore, cell viability was assessed by MTT assay. Results: HUVEC pretreatment with L-ser protected endothelial cells from H2O2 dependent cytotoxicity increased the surviving endothelial cell significantly. L-ser also led to concentration dependent increase of HO-1 (antioxidant defense protein) and Nrf2 (transcriptional factor that stimulate HO-1 gene expression) in cultured endothelial cells. Conclusion: These findings demonstrate that L-ser increased the level of HO-1 in endothelial cells. Increased HO-1 activity improves endothelial dysfunction suggesting an antiatherogenic potential of L-ser.
Poster — [A-10-1218-2] Pro-oxidant–antioxidant balance stature in thalassemia diseases Ghahramanlu Elhama, Ghayour Mobarhan Majidb, Tavallaie Shimac, Mirhossini Naghmed, Banihashem Abdollahd a Blood Transfusion Center, Islamic Republic of Iran b Ferdoosi University, Islamic Republic of Iran c Booali Research Center, Islamic Republic of Iran d Dr Sheikh Hospital, Islamic Republic of Iran E-mail addresses: [email protected] (G. Elham), [email protected] (G.M. Majid), [email protected] (T. Shima), [email protected] (M. Naghme)
Keywords: Antioxidant, L-serine, Heme oxygenase-1, HUVEC, Nrf2
doi:10.1016/j.clinbiochem.2011.08.940
Poster — [A-10-1207-1] Nucleostemin gene silencing by siRNA induces differentiation and growth inhibition of NB4 leukemia cell line Moosavi Mohammad Amin, Moghtaran Bonab Nazila, Hosseinpour Feizi Mohammad Ali Department of Zoology, Faculty of Natural Science, The University of Tabriz, Tabriz, Iran E-mail addresses: [email protected] (M.M. Amin), [email protected] (M.B. Nazila) Introduction: Acute promyelocytic leukemia (APL) is a malignant hematopoietic disorder that causes by indefinite proliferation and lack of differentiation of leukemia stem cells. Elucidation of signaling pathways involved in cancer stem cells is a current strategy in treatment of cancers. In this content, nucleostemin (NS), a nucleolar GTP binding protein, showed critical role in proliferation and apoptosis of cancer stem cells. Here, effects of NS gene silencing in NB4 cell line as an APL model have been studied for the first time. Methods: NB4 cell line was cultured in the presence of 200 nM NSsiRNA. NS gene expression levels were determined by RT-PCR. Trypan blue exclusion test was used to study growth inhibitory effects. Differentiation was studied morphologically and biologically using Wright–Giemsa staining and latex particle phagocytic activity, respectively. Results: The results showed that NS gene was highly expressed in NB4 cells. After 12–72 h of treatment with 200 nM siRNA, growth was inhibited up to 15–33%, respectively. After longer treatment times (48 h) differentiation characteristics such as decrease in nuclear cytoplasm ratio and ingestion of latex particles were observed, and confirmed the differentiation toward monocyte/macrophage.
Keywords: Nucleostemin, NB4, Differentiation, Growth inhibition doi:10.1016/j.clinbiochem.2011.08.941
Introduction: In beta-thalassemia major excess presence of the alpha globin chains is a primary reason for the cellular oxidative damage and also iron overload. The increased production of ROS might be associated with increased hemolysis in beta thalassemia major patients. Therefore, we wish to measure PAB as a marker of oxidative stress in thalassemia patients and healthy subjects. Purpose: To know that disturbances of pro-oxidant–antioxidant balance (PAB) occur in thalassemia diseases. Method: Prooxidant and antioxidant balance status was studied in 140 thalassemia major patients (8–20 years old ) in Sarve center, Mashhad, Iran, and their status was compared with 140 sex and agematched healthy subjects. Result: PAB value was significant higher in thalassemia patents than control group (78.41 vs.70.70). Conclusion: Thalassemic patients had high oxidative stress and these thalassemic children should improve oxidative stress by dietary advice. Also, PAB may be used as a simple measurement of oxidative stress (iron toxicity ) in thalasemic major patients. Keywords: PAB, Thalassemia, Oxidative stress doi:10.1016/j.clinbiochem.2011.08.942
Poster — [A-10-1218-3] Anti HSP 27 stature in thalassemic patients compared with healthy subjects Ghayour-Mobarhan Majida, Ghahramanlu Elhamb, Tavallaie Shimac, Abbaspur Alirezad, Mirhossini Naghmed, Banihashem Abdollahe a Biochemical and Nutritional Research Centre, Mashhad University of Medical Sciences, Mashhad, Iran b Blood Transfusion Center, Iran c Booali Research Center, Iran d Bojnourd University, Iran e Dr Sheikh Hospital, Iran E-mail addresses: [email protected] (G.-M. Majid), [email protected] (G. Elham), [email protected] (T. Shima), [email protected] (A. Alireza), [email protected] (M. Naghme) Introduction: HSP27 appears to have a dynamic and diverse range of functions in cellular survival. Heat shock proteins appear to
Abstracts
a
Department of Clinical Biochemistry, Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Hezarjarib Ave, Isfahan, Iran b Applied Physiology Research Center, Department of Physiology, Isfahan University of Medical Sciences, Isfahan, Iran E-mail addresses: [email protected] (N.M. Masoud), [email protected] (M. Ahmad), [email protected] (H.J. Shaghayegh)
Conclusion: Based on differentiating and growth inhibitory effects of NS-siRNA, NS can be proposed as a new target for treatment of APL patients.
Introduction: Oxidative stress is a causative factor of endothelial dysfunction and plays an important role in the pathophysiology of several vascular diseases. L-serine (L-ser) is known to exert cytoprotection in hepatocytes against oxidative stress. The present study investigates the effect of L-ser on H2O2-mediated cytotoxicity in cultured human umbilical vien endothelial cells (HUVEC). Materials and methods: In this study HUVEC were treated with different concentrations of L-Ser (0.1-3.2 mM). Then, Heme Oxygenase1(HO-1) and Nuclear factor-erythroid 2-related factor 2(Nrf2) levels were evaluated through Sandwich ELISA and DNA-binding based techniques respectively. Furthermore, cell viability was assessed by MTT assay. Results: HUVEC pretreatment with L-ser protected endothelial cells from H2O2 dependent cytotoxicity increased the surviving endothelial cell significantly. L-ser also led to concentration dependent increase of HO-1 (antioxidant defense protein) and Nrf2 (transcriptional factor that stimulate HO-1 gene expression) in cultured endothelial cells. Conclusion: These findings demonstrate that L-ser increased the level of HO-1 in endothelial cells. Increased HO-1 activity improves endothelial dysfunction suggesting an antiatherogenic potential of L-ser.
Poster — [A-10-1218-2] Pro-oxidant–antioxidant balance stature in thalassemia diseases Ghahramanlu Elhama, Ghayour Mobarhan Majidb, Tavallaie Shimac, Mirhossini Naghmed, Banihashem Abdollahd a Blood Transfusion Center, Islamic Republic of Iran b Ferdoosi University, Islamic Republic of Iran c Booali Research Center, Islamic Republic of Iran d Dr Sheikh Hospital, Islamic Republic of Iran E-mail addresses: [email protected] (G. Elham), [email protected] (G.M. Majid), [email protected] (T. Shima), [email protected] (M. Naghme)
Keywords: Antioxidant, L-serine, Heme oxygenase-1, HUVEC, Nrf2
doi:10.1016/j.clinbiochem.2011.08.940
Poster — [A-10-1207-1] Nucleostemin gene silencing by siRNA induces differentiation and growth inhibition of NB4 leukemia cell line Moosavi Mohammad Amin, Moghtaran Bonab Nazila, Hosseinpour Feizi Mohammad Ali Department of Zoology, Faculty of Natural Science, The University of Tabriz, Tabriz, Iran E-mail addresses: [email protected] (M.M. Amin), [email protected] (M.B. Nazila) Introduction: Acute promyelocytic leukemia (APL) is a malignant hematopoietic disorder that causes by indefinite proliferation and lack of differentiation of leukemia stem cells. Elucidation of signaling pathways involved in cancer stem cells is a current strategy in treatment of cancers. In this content, nucleostemin (NS), a nucleolar GTP binding protein, showed critical role in proliferation and apoptosis of cancer stem cells. Here, effects of NS gene silencing in NB4 cell line as an APL model have been studied for the first time. Methods: NB4 cell line was cultured in the presence of 200 nM NSsiRNA. NS gene expression levels were determined by RT-PCR. Trypan blue exclusion test was used to study growth inhibitory effects. Differentiation was studied morphologically and biologically using Wright–Giemsa staining and latex particle phagocytic activity, respectively. Results: The results showed that NS gene was highly expressed in NB4 cells. After 12–72 h of treatment with 200 nM siRNA, growth was inhibited up to 15–33%, respectively. After longer treatment times (48 h) differentiation characteristics such as decrease in nuclear cytoplasm ratio and ingestion of latex particles were observed, and confirmed the differentiation toward monocyte/macrophage.
Keywords: Nucleostemin, NB4, Differentiation, Growth inhibition doi:10.1016/j.clinbiochem.2011.08.941
Introduction: In beta-thalassemia major excess presence of the alpha globin chains is a primary reason for the cellular oxidative damage and also iron overload. The increased production of ROS might be associated with increased hemolysis in beta thalassemia major patients. Therefore, we wish to measure PAB as a marker of oxidative stress in thalassemia patients and healthy subjects. Purpose: To know that disturbances of pro-oxidant–antioxidant balance (PAB) occur in thalassemia diseases. Method: Prooxidant and antioxidant balance status was studied in 140 thalassemia major patients (8–20 years old ) in Sarve center, Mashhad, Iran, and their status was compared with 140 sex and agematched healthy subjects. Result: PAB value was significant higher in thalassemia patents than control group (78.41 vs.70.70). Conclusion: Thalassemic patients had high oxidative stress and these thalassemic children should improve oxidative stress by dietary advice. Also, PAB may be used as a simple measurement of oxidative stress (iron toxicity ) in thalasemic major patients. Keywords: PAB, Thalassemia, Oxidative stress doi:10.1016/j.clinbiochem.2011.08.942
Poster — [A-10-1218-3] Anti HSP 27 stature in thalassemic patients compared with healthy subjects Ghayour-Mobarhan Majida, Ghahramanlu Elhamb, Tavallaie Shimac, Abbaspur Alirezad, Mirhossini Naghmed, Banihashem Abdollahe a Biochemical and Nutritional Research Centre, Mashhad University of Medical Sciences, Mashhad, Iran b Blood Transfusion Center, Iran c Booali Research Center, Iran d Bojnourd University, Iran e Dr Sheikh Hospital, Iran E-mail addresses: [email protected] (G.-M. Majid), [email protected] (G. Elham), [email protected] (T. Shima), [email protected] (A. Alireza), [email protected] (M. Naghme) Introduction: HSP27 appears to have a dynamic and diverse range of functions in cellular survival. Heat shock proteins appear to