- Email: [email protected]
Expression of angiogenic marker CD34 in gastric adenocarcinoma and its relationship with cliniclopathological characteristics
S74
Abstracts
to E2 family of enzyme. We have previously characterized a homologue of this gene, UBE2Q2 (15q23), and reported that it is over-expressed in head and neck squamous cell carcinoma and breast cancer. We extended our studies to UBE2Q2 gene and its expression in some malignancies including ALL. In the present study the expression of this gene in leukemia was investigated. Methods: A series of RT-PCR and Quantitative Real-Time PCR was performed on a collection of 20 bone marrow samples of ALL patients and 20 whole blood samples of normal children. Then the relative quantification of our target gene transcripts was compared with reference gene (RPLP0 gene) transcript. Results: According to the results of RT-PCR, 90% of normal cells expressed UBE2Q1 gene while this amount is 45% for leukemic samples. Data obtained from QRT-PCR was in agreement with RT-PCR results and revealed that in 75% of the cases the gene is down regulated (2–3 fold). Conclusion: Our data suggest that this novel human gene, may have implications for pathogenesis of ALL and could be used in molecular diagnosis purposes and drug targets for treatment in the future. Keywords: E2 enzyme, UBE2Q1, New human gene, Acute lymphoblastic leukemia, QRT-PCR, RT-PCR doi:10.1016/j.clinbiochem.2011.08.161
E Poster – [A-10-972-3] An overview through p53 structure, function and regulation Fiuji Hamida,e, Ostadi Hassanb, Nassiry Mohammad Rezac, Mirhafez Rezad, Ghovati Shahrokhc a Molecular Cell Biology Department, FALW, VU University Amsterdam, Amsterdam, The Netherlands b Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran c Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran d Molecular Medicine, Department of Modern Sciences and Technologies, Faculty of Medicine, Mashhad University of Medical Science, Mashhad, Iran e Biochemistry Department, Payam Nour University, Mashhad, Iran E-mail address: [email protected] (F. Hamid) P53 tumor suppressor is a transcription factor with a very high turnover rate involving in cell cycle regulation, apoptosis and DNA repair. The system in which p53 plays role is activated by cellular stress such as DNA damage. Signals are transferred to the p53 via other proteins. The result is binding the p53 to the target promoter regions on DNA to activate their transcription. The products of target genes can induct DNA repair mechanism or programmed cell death. In most of the cancers, about 50%, inactive p53 has been detected. Malfunction of the p53 is due to various mutations that can make changes in p53 structure, stability or its different abilities. Understanding the molecular structure of p53 provides ways to find pathways of therapeutic reactivation of p53 favors apoptotic cell death in tumor cells. Keywords: p53, Mdm2, Cancer therapy, Apoptosis, p53 regulation doi:10.1016/j.clinbiochem.2011.08.162
Oral – [A-10-986-1] Construction of a recombinant Leishmania tarentolae expressing human papillomavirus type 16 E7 gene and evaluation of its immunogenicity in C57BL/6 mice model Maryam Salehiab, Azam Bolhassanib, Tahereh Taherib, Elham Mohitb, Negar Seyedb, Farnaz Zahedifardb, Yasaman Taslimib, Mandana Sattariab, Sima Rafatib
a
Velenjak, Shahid Beheshti University of Medical Sciences, Iran Pasteur Institute of Iran, Tehran, Iran E-mail addresses: [email protected] (M. Salehi), [email protected] (A. Bolhassani), [email protected] (T. Taheri), [email protected] (E. Mohit), [email protected] (N. Seyed), [email protected] (F. Zahedifard), [email protected] (Y. Taslimi), [email protected] (M. Sattari), [email protected] (S. Rafati)
b
Introduction: The second largest cause of cancer deaths in women, cervical cancer, is associated with human papillomaviruses (HPVs). E7 is the key oncogenic protein in the induction of HPV-related malignancies. Thus, producing an efficient vaccine against HPV by targeting E7 protein can be a promising strategy in HPV vaccination. Among various vaccination approaches against cervical cancer, the recent live vaccine strategies using non pathogenic Leishmania tarentolae could be another approach toward vaccine development. Methods: In order to design a live vaccine against HPV infection, we stably expressed E7-GFP fusion gene in a non-pathogenic lizard protozoan, L. tarentolae, using homologous recombination. Western blot analysis was used for fusion protein confirmation. Large-scale purification of a plasmid DNA sample (pcDNA-E7) was performed by an endotoxin free kit. Then, humoral and cellular immune responses were evaluated in different groups as a live/live vaccine strategy in comparison with two other approaches including DNA/DNA and DNA/ live in a C57BL/6 mice model. Results: The E7-GFP protein expression in recombinant L. tarentolae was confirmed by western blotting. Our results indicated that subcutaneous administration of the E7-GFP recombinant L. tarentolae strain induces both IgG2a and IFN-γ as compared to other strategies. Furthermore, there were no significant differences for IL-5 production in different groups as compared with that in control groups. Conclusion: Our data shows that live vaccine is successful in eliciting the proper immune response. This study likely provides an additional option for inducing efficient immune responses as a possible immune therapy modality against HPV infections. Keywords: HPV, Cancer, E7, Live vaccine, Leishmania tarentolae doi:10.1016/j.clinbiochem.2011.08.163
E Poster – [A-10-1009-3] Expression of angiogenic marker CD34 in gastric adenocarcinoma and its relationship with cliniclopathological characteristics Ghaffarzadehgan Kamrana, Ghadri Sepidehb, Raziee Hamid Rezac, Hosseinnezhad Haniehd, Moaven Omeedc, Abdollahi Tahoorac, Sima Hamid Rezac a Department of Pathology, Razavi Hospital, Mashhad, Iran b Imam Reza Hospital, Mashhad University of Medical Sciences, Mashhad, Iran c Department of Medicine, Imam Reza Hospital, Mashhad University of Medical Sciences, Iran d Mashhad, Malek abad Ave, Nastaren 18, No 113, 3th stair, Iran E-mail addresses: [email protected] (G. Kamran), [email protected] (G. Sepideh), [email protected] (R.H. Reza), [email protected] (H. Hanieh), [email protected] (M. Omeed), [email protected] (S.H. Reza) Introduction: Gastric cancer (GC) is still one of the most leading causes of cancer-related death worldwide. GC considered as a multistage progressive process in which tumor growth and metastasis spread
Abstracts
mainly caused by angiogenesis process. CD34 is a glycoprotein which present in the vascular endothelium is a new biomarker for the evaluation of angiogenesis quantification and tumor development. Materials and methods: The aim of this study is to measure the immunohistochemical expression of CD34 in gastric adenocarcinoma tissue to identify the prognostic value and its relationship with the clinicopathological parameters. Samples obtained from 29 gastric adenocarcinoma patients who had undergone gastrectomy. 5 fields of samples were counted, and the median of them were reported. The range up to 25% reported as a high positivity expression of CD34. Results: Based on our data all of 29 cases registered in our study were CD34 positive. The statistical analysis showed that, there is no significant relationship between age, duration of problem, tumor stage and grade with high/low positivity expression of CD34. Conclusion: Our analyses showed a significant correlation between CD34 tumor tissue expression and tumor location. Cardia-tumors significantly showed a highly positivity expression of CD34 (81.8%) VS non cardia-tumors (24%). Keywords: Angiogenic marker, CD34, Gastric adenocarcinoma, Cliniclopathologic feature doi:10.1016/j.clinbiochem.2011.08.164
E Poster – [A-10-1126-2] Down-regulation of IAP proteins concomitant with the activation of Caspase-3 and -9 by a derivative from 4-aryl-4H-chromenes family induces apoptosis in HepG2 cancer cell line Mahdavi Majid, Davoodi Jamshid, Mohebbi Seyed Reza, Zali Mohammad Reza, Foroumadi Alireza Institute of Biochemistry and Biophysics, the University of Tehran, Tehran, Iran E-mail addresses: [email protected] (M. Majid), [email protected] (D. Jamshid) Introduction: Since many cancerous cells exhibit abnormal inhibition of apoptosis, researchers are interested in the discovery and development of apoptotic inducers which serve as potential anticancer agents. Discovery of compounds belonging to the 4-aryl4H-chromenes family with anti-cancer activity has been recently reported. Here we reported a derivative of this family with high apoptotic activity against liver cancer HepG2. Methods: HepG2 cells were seeded in 24-well plates at 1 × 10 cells/well and treated with 50–80 nM of 2-amino-4-(3-nitro phenyl)3-cyano-7-(dimethylamino)-4Hchromene (3-NC). This compound was found to be highly active growth inhibitor with IC50 values of 55 ± 3.2 nM as determined by MTT assay and cell viability decreased more than 50% upon 72 hours of treatment at 50–80 nM concentration of the compound. Apoptosis, as a kind of programmed cell death was investigated morphologically by Hoechst 33258 staining, caspase-3/7 like activation assay, as well as formation of DNA ladder. Result: Treatment of HepG2 cells with 3-NC (at IC50 Value) indicated that more than 50% of HepG2 cells underwent apoptosis and also caspase 3/7 activity increased by more than 40% following 72 h treatment. In addition to increased caspase-3/7 like activity, caspase-9 was also cleaved indicating the activation of the intrinsic apoptosis pathway. Furthermore, Western blot analysis showed 3NC is also able to down-regulate the expression of IAPs, survivin, cIAP2 and XIAP, anti-apoptotic proteins that are highly expressed in tumor cells. Conclusion: The results of this investigation clearly indicated that 3-NC is capable of inducing apoptosis in HepG2 cells and therefore, could be a valuable candidate for pharmeceutical evaluations.
S75
Keywords: Apoptosis, 4-aryl-4H-chromenes, Caspase-3, Caspase-9, cIAP2, Survivin, XIAP doi:10.1016/j.clinbiochem.2011.08.165
Nano–Biotechnology: Applications in Biochemistry Poster – [A-10-77-5] Survey characterization nano structure surface layer in some of pathogen bacteria Jalalpoor Shilla Islamic Azad University Shahreza Branch, Isfahan, Iran E-mail address: [email protected] Introduction: An S-layer (surface layer) is a part of the cell envelope commonly found in bacteria, as well as among archaea. It consists of a monomolecular layer composed of identical proteins or glycoproteins. As for many bacteria the S-layer represents the outermost interaction zone with their respective environment, its functions are very diverse and vary from species to species. Methods: Related papers to surface layer were extracted from articles in Pubmed, Elsevier Science, and Yahoo from years 1995 to 2010. For this study key words which were searched include surface layer, pathogenesis, and pathogen bacteria. Results: About the surface layer in all of similar original and review articles, there is consensus that the existence of this surface structure in bacteria lead to increased pathogenesity in bacteria. Conclusion: S-layers in pathogen bacteria protection against bacteriophages and phagocytosis, resistance against low pH, adhesion, stabilisation of the membrane and provide adhesion sites for exoproteins due to more pathogenesis, infection resistant and antibiotic resistant in bacteria. Result of this search showed prevalent S-layer in pathogen bacteria and importantly determined S-layer producer strains in laboratory. Keywords: Nano structure surface layer, Pathogen bacteria, Archaea, Pathogenesis doi:10.1016/j.clinbiochem.2011.08.166
Poster – [A-10-77-6] Survey frequency of virulent agent in Bacillus cereus strains Jalalpoor Shilla Islamic Azad University, Shahreza Branch, Shahreza, Iran E-mail address: [email protected] Introduction: S-layer is the outermost protein in bacteria, S-layer inhibits antibiotics in bacteria and due to increase of pathogenicity in bacteria, according to role of staff hands and hospital surfaces in nosocomial infections, therefore contamination of this source with Bacillus cereus strain producer S-layer and β-lactamase, due to spread of antibiotic resistant nosocomial infections, S-layer and β–lactamase are important virulent agent in B. cereus. Objective: This search was a survey of the frequency of S-layer and β-lactamase and the role of S-layer in antibiotics to inhibit in B. cereus strain isolated of staff hands and hospital surfaces. Methods: The research was performed with laboratory method during the years 2005/2007 in Azzahra hospital and Isfahan University, according to statistical formula study of 274 samples. For preparation of samples, cultured bacteria in TSA and specimen's electrophoresis with 10X SDS-PAGE. Sample antibiogram was performed with Kirby Bauer method and β-lactamase production, with acidimetric method. All the statistical analyses are carried out using SPSS version 14. Chi-
Abstracts
to E2 family of enzyme. We have previously characterized a homologue of this gene, UBE2Q2 (15q23), and reported that it is over-expressed in head and neck squamous cell carcinoma and breast cancer. We extended our studies to UBE2Q2 gene and its expression in some malignancies including ALL. In the present study the expression of this gene in leukemia was investigated. Methods: A series of RT-PCR and Quantitative Real-Time PCR was performed on a collection of 20 bone marrow samples of ALL patients and 20 whole blood samples of normal children. Then the relative quantification of our target gene transcripts was compared with reference gene (RPLP0 gene) transcript. Results: According to the results of RT-PCR, 90% of normal cells expressed UBE2Q1 gene while this amount is 45% for leukemic samples. Data obtained from QRT-PCR was in agreement with RT-PCR results and revealed that in 75% of the cases the gene is down regulated (2–3 fold). Conclusion: Our data suggest that this novel human gene, may have implications for pathogenesis of ALL and could be used in molecular diagnosis purposes and drug targets for treatment in the future. Keywords: E2 enzyme, UBE2Q1, New human gene, Acute lymphoblastic leukemia, QRT-PCR, RT-PCR doi:10.1016/j.clinbiochem.2011.08.161
E Poster – [A-10-972-3] An overview through p53 structure, function and regulation Fiuji Hamida,e, Ostadi Hassanb, Nassiry Mohammad Rezac, Mirhafez Rezad, Ghovati Shahrokhc a Molecular Cell Biology Department, FALW, VU University Amsterdam, Amsterdam, The Netherlands b Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran c Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran d Molecular Medicine, Department of Modern Sciences and Technologies, Faculty of Medicine, Mashhad University of Medical Science, Mashhad, Iran e Biochemistry Department, Payam Nour University, Mashhad, Iran E-mail address: [email protected] (F. Hamid) P53 tumor suppressor is a transcription factor with a very high turnover rate involving in cell cycle regulation, apoptosis and DNA repair. The system in which p53 plays role is activated by cellular stress such as DNA damage. Signals are transferred to the p53 via other proteins. The result is binding the p53 to the target promoter regions on DNA to activate their transcription. The products of target genes can induct DNA repair mechanism or programmed cell death. In most of the cancers, about 50%, inactive p53 has been detected. Malfunction of the p53 is due to various mutations that can make changes in p53 structure, stability or its different abilities. Understanding the molecular structure of p53 provides ways to find pathways of therapeutic reactivation of p53 favors apoptotic cell death in tumor cells. Keywords: p53, Mdm2, Cancer therapy, Apoptosis, p53 regulation doi:10.1016/j.clinbiochem.2011.08.162
Oral – [A-10-986-1] Construction of a recombinant Leishmania tarentolae expressing human papillomavirus type 16 E7 gene and evaluation of its immunogenicity in C57BL/6 mice model Maryam Salehiab, Azam Bolhassanib, Tahereh Taherib, Elham Mohitb, Negar Seyedb, Farnaz Zahedifardb, Yasaman Taslimib, Mandana Sattariab, Sima Rafatib
a
Velenjak, Shahid Beheshti University of Medical Sciences, Iran Pasteur Institute of Iran, Tehran, Iran E-mail addresses: [email protected] (M. Salehi), [email protected] (A. Bolhassani), [email protected] (T. Taheri), [email protected] (E. Mohit), [email protected] (N. Seyed), [email protected] (F. Zahedifard), [email protected] (Y. Taslimi), [email protected] (M. Sattari), [email protected] (S. Rafati)
b
Introduction: The second largest cause of cancer deaths in women, cervical cancer, is associated with human papillomaviruses (HPVs). E7 is the key oncogenic protein in the induction of HPV-related malignancies. Thus, producing an efficient vaccine against HPV by targeting E7 protein can be a promising strategy in HPV vaccination. Among various vaccination approaches against cervical cancer, the recent live vaccine strategies using non pathogenic Leishmania tarentolae could be another approach toward vaccine development. Methods: In order to design a live vaccine against HPV infection, we stably expressed E7-GFP fusion gene in a non-pathogenic lizard protozoan, L. tarentolae, using homologous recombination. Western blot analysis was used for fusion protein confirmation. Large-scale purification of a plasmid DNA sample (pcDNA-E7) was performed by an endotoxin free kit. Then, humoral and cellular immune responses were evaluated in different groups as a live/live vaccine strategy in comparison with two other approaches including DNA/DNA and DNA/ live in a C57BL/6 mice model. Results: The E7-GFP protein expression in recombinant L. tarentolae was confirmed by western blotting. Our results indicated that subcutaneous administration of the E7-GFP recombinant L. tarentolae strain induces both IgG2a and IFN-γ as compared to other strategies. Furthermore, there were no significant differences for IL-5 production in different groups as compared with that in control groups. Conclusion: Our data shows that live vaccine is successful in eliciting the proper immune response. This study likely provides an additional option for inducing efficient immune responses as a possible immune therapy modality against HPV infections. Keywords: HPV, Cancer, E7, Live vaccine, Leishmania tarentolae doi:10.1016/j.clinbiochem.2011.08.163
E Poster – [A-10-1009-3] Expression of angiogenic marker CD34 in gastric adenocarcinoma and its relationship with cliniclopathological characteristics Ghaffarzadehgan Kamrana, Ghadri Sepidehb, Raziee Hamid Rezac, Hosseinnezhad Haniehd, Moaven Omeedc, Abdollahi Tahoorac, Sima Hamid Rezac a Department of Pathology, Razavi Hospital, Mashhad, Iran b Imam Reza Hospital, Mashhad University of Medical Sciences, Mashhad, Iran c Department of Medicine, Imam Reza Hospital, Mashhad University of Medical Sciences, Iran d Mashhad, Malek abad Ave, Nastaren 18, No 113, 3th stair, Iran E-mail addresses: [email protected] (G. Kamran), [email protected] (G. Sepideh), [email protected] (R.H. Reza), [email protected] (H. Hanieh), [email protected] (M. Omeed), [email protected] (S.H. Reza) Introduction: Gastric cancer (GC) is still one of the most leading causes of cancer-related death worldwide. GC considered as a multistage progressive process in which tumor growth and metastasis spread
Abstracts
mainly caused by angiogenesis process. CD34 is a glycoprotein which present in the vascular endothelium is a new biomarker for the evaluation of angiogenesis quantification and tumor development. Materials and methods: The aim of this study is to measure the immunohistochemical expression of CD34 in gastric adenocarcinoma tissue to identify the prognostic value and its relationship with the clinicopathological parameters. Samples obtained from 29 gastric adenocarcinoma patients who had undergone gastrectomy. 5 fields of samples were counted, and the median of them were reported. The range up to 25% reported as a high positivity expression of CD34. Results: Based on our data all of 29 cases registered in our study were CD34 positive. The statistical analysis showed that, there is no significant relationship between age, duration of problem, tumor stage and grade with high/low positivity expression of CD34. Conclusion: Our analyses showed a significant correlation between CD34 tumor tissue expression and tumor location. Cardia-tumors significantly showed a highly positivity expression of CD34 (81.8%) VS non cardia-tumors (24%). Keywords: Angiogenic marker, CD34, Gastric adenocarcinoma, Cliniclopathologic feature doi:10.1016/j.clinbiochem.2011.08.164
E Poster – [A-10-1126-2] Down-regulation of IAP proteins concomitant with the activation of Caspase-3 and -9 by a derivative from 4-aryl-4H-chromenes family induces apoptosis in HepG2 cancer cell line Mahdavi Majid, Davoodi Jamshid, Mohebbi Seyed Reza, Zali Mohammad Reza, Foroumadi Alireza Institute of Biochemistry and Biophysics, the University of Tehran, Tehran, Iran E-mail addresses: [email protected] (M. Majid), [email protected] (D. Jamshid) Introduction: Since many cancerous cells exhibit abnormal inhibition of apoptosis, researchers are interested in the discovery and development of apoptotic inducers which serve as potential anticancer agents. Discovery of compounds belonging to the 4-aryl4H-chromenes family with anti-cancer activity has been recently reported. Here we reported a derivative of this family with high apoptotic activity against liver cancer HepG2. Methods: HepG2 cells were seeded in 24-well plates at 1 × 10 cells/well and treated with 50–80 nM of 2-amino-4-(3-nitro phenyl)3-cyano-7-(dimethylamino)-4Hchromene (3-NC). This compound was found to be highly active growth inhibitor with IC50 values of 55 ± 3.2 nM as determined by MTT assay and cell viability decreased more than 50% upon 72 hours of treatment at 50–80 nM concentration of the compound. Apoptosis, as a kind of programmed cell death was investigated morphologically by Hoechst 33258 staining, caspase-3/7 like activation assay, as well as formation of DNA ladder. Result: Treatment of HepG2 cells with 3-NC (at IC50 Value) indicated that more than 50% of HepG2 cells underwent apoptosis and also caspase 3/7 activity increased by more than 40% following 72 h treatment. In addition to increased caspase-3/7 like activity, caspase-9 was also cleaved indicating the activation of the intrinsic apoptosis pathway. Furthermore, Western blot analysis showed 3NC is also able to down-regulate the expression of IAPs, survivin, cIAP2 and XIAP, anti-apoptotic proteins that are highly expressed in tumor cells. Conclusion: The results of this investigation clearly indicated that 3-NC is capable of inducing apoptosis in HepG2 cells and therefore, could be a valuable candidate for pharmeceutical evaluations.
S75
Keywords: Apoptosis, 4-aryl-4H-chromenes, Caspase-3, Caspase-9, cIAP2, Survivin, XIAP doi:10.1016/j.clinbiochem.2011.08.165
Nano–Biotechnology: Applications in Biochemistry Poster – [A-10-77-5] Survey characterization nano structure surface layer in some of pathogen bacteria Jalalpoor Shilla Islamic Azad University Shahreza Branch, Isfahan, Iran E-mail address: [email protected] Introduction: An S-layer (surface layer) is a part of the cell envelope commonly found in bacteria, as well as among archaea. It consists of a monomolecular layer composed of identical proteins or glycoproteins. As for many bacteria the S-layer represents the outermost interaction zone with their respective environment, its functions are very diverse and vary from species to species. Methods: Related papers to surface layer were extracted from articles in Pubmed, Elsevier Science, and Yahoo from years 1995 to 2010. For this study key words which were searched include surface layer, pathogenesis, and pathogen bacteria. Results: About the surface layer in all of similar original and review articles, there is consensus that the existence of this surface structure in bacteria lead to increased pathogenesity in bacteria. Conclusion: S-layers in pathogen bacteria protection against bacteriophages and phagocytosis, resistance against low pH, adhesion, stabilisation of the membrane and provide adhesion sites for exoproteins due to more pathogenesis, infection resistant and antibiotic resistant in bacteria. Result of this search showed prevalent S-layer in pathogen bacteria and importantly determined S-layer producer strains in laboratory. Keywords: Nano structure surface layer, Pathogen bacteria, Archaea, Pathogenesis doi:10.1016/j.clinbiochem.2011.08.166
Poster – [A-10-77-6] Survey frequency of virulent agent in Bacillus cereus strains Jalalpoor Shilla Islamic Azad University, Shahreza Branch, Shahreza, Iran E-mail address: [email protected] Introduction: S-layer is the outermost protein in bacteria, S-layer inhibits antibiotics in bacteria and due to increase of pathogenicity in bacteria, according to role of staff hands and hospital surfaces in nosocomial infections, therefore contamination of this source with Bacillus cereus strain producer S-layer and β-lactamase, due to spread of antibiotic resistant nosocomial infections, S-layer and β–lactamase are important virulent agent in B. cereus. Objective: This search was a survey of the frequency of S-layer and β-lactamase and the role of S-layer in antibiotics to inhibit in B. cereus strain isolated of staff hands and hospital surfaces. Methods: The research was performed with laboratory method during the years 2005/2007 in Azzahra hospital and Isfahan University, according to statistical formula study of 274 samples. For preparation of samples, cultured bacteria in TSA and specimen's electrophoresis with 10X SDS-PAGE. Sample antibiogram was performed with Kirby Bauer method and β-lactamase production, with acidimetric method. All the statistical analyses are carried out using SPSS version 14. Chi-