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Investigation on the relation of CDC25A gene and breast cancer
S204
Abstracts
Poster – [A-10-877-1] Molecular analysis of rs1799777 polymorphism of MLH1 gene in colorectal cancer susceptibility Milanizadeh Saman, Montazer Haghighi Mahdi, Damavand Behzad, Majidizadeh Bozorgi Sajad, Khanyaghma Mahsa, Mirtalebi Hanye, Fatemi Seyyed Reza, Zali Mohammadreza Taleghani hospital, Iran E-mail addresses: [email protected] (M. Saman), [email protected] (M.H. Mahdi), [email protected] (D. Behzad), [email protected] (M.B. Sajad), [email protected] (K. Mahsa), [email protected] (M. Hanye), [email protected] (F.S. Reza), [email protected] (Z. Mohammadreza) Introduction: Recent National Cancer Registration reports show that colorectal cancer is the third common cancer in the women and fifth among men in Iranian population. Seventy percent of colorectal cancer encountered patients are sporadic. One possible way for cancer affection is alteration in MMR system genes. Thus, analysis of efficient genes such as MLH1 has a critical role in diagnosis and prognosis of this cancer. Materials and methods: 140 patients with sporadic colorectal cancer and 135 control subjects were selected for this analysis. DNA was extracted with salting-out method. Polymorph region of MLH1 gene was amplified with PCR and genotyping was carried out by RFLP. 10% of PCR products were genotyped by direct sequencing. Results: This study concluded that A allele frequency was 0.7 and G allele frequency was 0.3. We found that A/G genotype of rs1799777 increases colorectal cancer susceptibility around 6 fold (P < 0.005, CI95% = 12.543–3.236 and OR = 6.142). But there aren't any significant relation between other genotypes and colorectal cancer. Conclusion: These results confirm previous studies in other populations on rs179977 about relation of this SNP with colorectal cancer, consequently, prove importance of this SNP and MLH1 gene on colorectal cancer susceptibility. Keywords: Colorectal cancer, MMR, MLH1, Polymorphism
doi:10.1016/j.clinbiochem.2011.08.503
E.Poster – [A-10-924-1] Investigation on the relation of CDC25A gene and breast cancer Leili Khakpoura, Majid Motovali Bashib, Shirin Farivarc a Department of Genetics, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran b Genetic division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran c Department of Genetics, Faculty of Biological Science, Shahid Beheshti University, Tehran, Iran E-mail addresses: [email protected] (L. Khakpour), [email protected] (M. Motovali Bashi), [email protected] (S. Farivar) Introduction: Breast cancer is the most common malignant tumor among women. It has been shown that breast cancer results from multiple environmental factors as well as genetic variations, such as genetic polymorphisms. The family of cell division cycle 25 (CDC25) phosphatase is one of the important regulators for cell cycle progression. CDC25A activates the cyclin-dependent kinase CDC2 by removing two phosphate groups. It is required for transition from G1 to the S phase. Recent studies have shown that this gene is overexpressed in breast tumors. CDC25A is located on 3p21 and includes 15 exons. This study aims to investigate the correlation
between the human 263 C/T polymorphism in CDC25A gene with breast cancer. Materials and Methods: DNA extraction was performed using blood samples which were collected from patients of breast cancer and controls. The primers were designed by NCBI gene bank and oligo-7 software. Sample genotyping was performed using PCR-RFLP technique. Results: PCR product bands on the gel electrophoresis were located exactly on the expected place. So far, PCR products of 40 controls and 50 cases were digested with restriction enzyme as expected; hence they were genotyped by the method. Conclusion: Our initial results show that this gene is associated with breast cancer. Significant differences between patients and controls can lead to finding a marker for earlier diagnosis and targets for cancer therapy. Keywords: Breast cancer, CDC25A, SNP, PCR-RFLP
doi:10.1016/j.clinbiochem.2011.08.504
Poster – [A-10-942-1] Study of the construction of an adenoviral vector for hepatic tumor gene therapy application Bahare Mehrdad Vahdati, Hossein A. Tehrani, Seyed Younes Hosseini Dept. of Medical Biotechnology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran E-mail addresses: [email protected] (B.M. Vahdati), [email protected] (H.A. Tehrani), [email protected] (S.Y. Hosseini) Introduction: Hepatocellular carcinoma is one of the most common cancers worldwide. Currently, there is no effective therapy to this cancer. A promising approach is using gene therapy strategies with the help of the widely used adenoviral vectors which is safe to human. In this project, a study was performed to develop an adenoviral construct containing a suicide gene under the control of a tumor specific promoter. Methods: The cDNA of Midkine (MK) promoter and tBid gene had been cloned in pcDNA3.1 vector in our lab. The MK-tBid fragment was excised from this vector and replaced with CMV promoter region of the Ad shuttle vector of pAd-CMV5. The resulting construct was then transformed into DH5α strains, to propagate it. The presence of the cloned fragment was confirmed by colony PCR reaction, diagnostic digestion with specific restriction enzymes and DNA sequencing. Results: The results of colony PCR, restriction enzyme and DNA sequencing analysis showed that the final recombinant Ad shuttle vector (pAd-MD-tBid) was constructed successfully. Conclusion: In this research, tBid is an apoptotic gene that is under the control of midkine (MK), a tumor-specific promoter which is overexpressed in a variety of cancer cells but its expression is low in normal cells. The pAd-MD-tBid is capable of homologous recombination with a backbone adenoviral vector in E. coli, to generate recombinant Adenoviruses that can be used for therapeutic applications. When transfected, it causes specific therapeutic effect on hepatic tumor cells in comparison with normal cells. Keywords: Hepatocellular carcinoma, Adenovirus, suicide gene therapy
doi:10.1016/j.clinbiochem.2011.08.505
Abstracts
Poster – [A-10-877-1] Molecular analysis of rs1799777 polymorphism of MLH1 gene in colorectal cancer susceptibility Milanizadeh Saman, Montazer Haghighi Mahdi, Damavand Behzad, Majidizadeh Bozorgi Sajad, Khanyaghma Mahsa, Mirtalebi Hanye, Fatemi Seyyed Reza, Zali Mohammadreza Taleghani hospital, Iran E-mail addresses: [email protected] (M. Saman), [email protected] (M.H. Mahdi), [email protected] (D. Behzad), [email protected] (M.B. Sajad), [email protected] (K. Mahsa), [email protected] (M. Hanye), [email protected] (F.S. Reza), [email protected] (Z. Mohammadreza) Introduction: Recent National Cancer Registration reports show that colorectal cancer is the third common cancer in the women and fifth among men in Iranian population. Seventy percent of colorectal cancer encountered patients are sporadic. One possible way for cancer affection is alteration in MMR system genes. Thus, analysis of efficient genes such as MLH1 has a critical role in diagnosis and prognosis of this cancer. Materials and methods: 140 patients with sporadic colorectal cancer and 135 control subjects were selected for this analysis. DNA was extracted with salting-out method. Polymorph region of MLH1 gene was amplified with PCR and genotyping was carried out by RFLP. 10% of PCR products were genotyped by direct sequencing. Results: This study concluded that A allele frequency was 0.7 and G allele frequency was 0.3. We found that A/G genotype of rs1799777 increases colorectal cancer susceptibility around 6 fold (P < 0.005, CI95% = 12.543–3.236 and OR = 6.142). But there aren't any significant relation between other genotypes and colorectal cancer. Conclusion: These results confirm previous studies in other populations on rs179977 about relation of this SNP with colorectal cancer, consequently, prove importance of this SNP and MLH1 gene on colorectal cancer susceptibility. Keywords: Colorectal cancer, MMR, MLH1, Polymorphism
doi:10.1016/j.clinbiochem.2011.08.503
E.Poster – [A-10-924-1] Investigation on the relation of CDC25A gene and breast cancer Leili Khakpoura, Majid Motovali Bashib, Shirin Farivarc a Department of Genetics, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran b Genetic division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran c Department of Genetics, Faculty of Biological Science, Shahid Beheshti University, Tehran, Iran E-mail addresses: [email protected] (L. Khakpour), [email protected] (M. Motovali Bashi), [email protected] (S. Farivar) Introduction: Breast cancer is the most common malignant tumor among women. It has been shown that breast cancer results from multiple environmental factors as well as genetic variations, such as genetic polymorphisms. The family of cell division cycle 25 (CDC25) phosphatase is one of the important regulators for cell cycle progression. CDC25A activates the cyclin-dependent kinase CDC2 by removing two phosphate groups. It is required for transition from G1 to the S phase. Recent studies have shown that this gene is overexpressed in breast tumors. CDC25A is located on 3p21 and includes 15 exons. This study aims to investigate the correlation
between the human 263 C/T polymorphism in CDC25A gene with breast cancer. Materials and Methods: DNA extraction was performed using blood samples which were collected from patients of breast cancer and controls. The primers were designed by NCBI gene bank and oligo-7 software. Sample genotyping was performed using PCR-RFLP technique. Results: PCR product bands on the gel electrophoresis were located exactly on the expected place. So far, PCR products of 40 controls and 50 cases were digested with restriction enzyme as expected; hence they were genotyped by the method. Conclusion: Our initial results show that this gene is associated with breast cancer. Significant differences between patients and controls can lead to finding a marker for earlier diagnosis and targets for cancer therapy. Keywords: Breast cancer, CDC25A, SNP, PCR-RFLP
doi:10.1016/j.clinbiochem.2011.08.504
Poster – [A-10-942-1] Study of the construction of an adenoviral vector for hepatic tumor gene therapy application Bahare Mehrdad Vahdati, Hossein A. Tehrani, Seyed Younes Hosseini Dept. of Medical Biotechnology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran E-mail addresses: [email protected] (B.M. Vahdati), [email protected] (H.A. Tehrani), [email protected] (S.Y. Hosseini) Introduction: Hepatocellular carcinoma is one of the most common cancers worldwide. Currently, there is no effective therapy to this cancer. A promising approach is using gene therapy strategies with the help of the widely used adenoviral vectors which is safe to human. In this project, a study was performed to develop an adenoviral construct containing a suicide gene under the control of a tumor specific promoter. Methods: The cDNA of Midkine (MK) promoter and tBid gene had been cloned in pcDNA3.1 vector in our lab. The MK-tBid fragment was excised from this vector and replaced with CMV promoter region of the Ad shuttle vector of pAd-CMV5. The resulting construct was then transformed into DH5α strains, to propagate it. The presence of the cloned fragment was confirmed by colony PCR reaction, diagnostic digestion with specific restriction enzymes and DNA sequencing. Results: The results of colony PCR, restriction enzyme and DNA sequencing analysis showed that the final recombinant Ad shuttle vector (pAd-MD-tBid) was constructed successfully. Conclusion: In this research, tBid is an apoptotic gene that is under the control of midkine (MK), a tumor-specific promoter which is overexpressed in a variety of cancer cells but its expression is low in normal cells. The pAd-MD-tBid is capable of homologous recombination with a backbone adenoviral vector in E. coli, to generate recombinant Adenoviruses that can be used for therapeutic applications. When transfected, it causes specific therapeutic effect on hepatic tumor cells in comparison with normal cells. Keywords: Hepatocellular carcinoma, Adenovirus, suicide gene therapy
doi:10.1016/j.clinbiochem.2011.08.505