Upregulated Myc Expression in N-Methyl Nitrosourea (MNU)-induced Rat Mammary Tumours

Barathidasan, Rajamani,Pawaiya, Rajveer Singh,Rai, Ram Bahal,Dhama, Kuldeep Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.8

Background: The most common incident cancer and cause of cancer-related deaths in women is breast cancer. The Myc gene is upregulated in many cancer types including breast cancer, and it is considered as a potential anti-cancer drug target. The present study was conducted to evaluate the Myc (gene and protein) expression pattern in an experimental mammary tumour model in rats. Materials and Methods: Thirty six Sprague Dawley rats were divided into: Experimental group (26 animals), which received the chemical carcinogen N-methyl nitrosourea (MNU) and a control group (10 animals), which received vehicle only. c-Myc oncoprotein and its mRNA expression pattern were evaluated using immunohistochemistry (IHC) and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), respectively, in normal rat mammary tissue and mammary tumours. The rat glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as internal control for semi-quantitative RT-PCR. Results: Histopathological examination of mammary tissues and tumours from MNU treated animals revealed the presence of premalignant lesions, benign tumours, in situ carcinomas and invasive carcinomas. Immunohistochemical evaluation of tumour tissues showed upregulation and heterogeneous cellular localization of c-Myc oncoprotein. The expression levels of c-Myc oncoprotein were significantly elevated (75-91%) in all the tumours. Semi-quantitative RT-PCR revealed increased expression of c-Myc mRNA in mammary tumours compared to normal mammary tissues. Conclusions: Further large-scale investigation study is needed to adopt this experimental rat mammary tumour model as an in vivo model to study anti-cancer strategies directed against Myc or its downstream partners at the transcriptional or post-transcriptional level.

Gene Expression Patterns of Spleen, Lung and Brain with Different Radiosensitivity in C57BL6 Mice

Majumder Md. Zahidur Rahman,Lee, Woo-Jung,Lee, Su-Jae,Bae, Sang-Woo,Lee, Yun-Sil The Korean Association for Radiation Protection 2005 방사선방어학회지 Vol.30 No.4

Although little information is available on the underlying mechanisms, various genetic factors have been associated with tissue-specific responses to radiation. In the present study, we explored the possibility whether organ specific gene expression is associated with radiosensitivity using samples from brain, lung and spleen. We examined intrinsic expression pattern of 23 genes in the organs by semi-quantitative RT-PCR method using both male and female C57BL/6 mice. Expression of p53 and p21, well known factors for governing sensitivity to radiation or chemotherapeutic agents, was not different among the organ types. Both higher expression of sialyltransferase, delta7-sterol reductase, leptin receptor splice variant form 12.1, and Cu/Zn superoxide dismutase (SOD) and lower expression of alphaB crystalline were specific for spleen tissue. Expression level of glutathione peroxidase and APO-1 cell surface antigen gene in lung tissue was high, while that of Na, K-ATPase alpha-subunit, Cu/ZnSOD, and cyclin G was low. Brain, radioresistant organ, showed higher expressions of Na, K-ATPase-subunit, cyclin G, and nucleolar protein hNop56 and lower expression of delta7-sterol reductase. The result revealed a potential correlation between gene expression patterns and organ sensitivity, and Identified genes which might be responsible for organ sensitivity.

Transcriptome Analysis of the Barley-Rhynchosporium secalis Interaction

Al-Daoude, Antonious,Shoaib, Amina,Al-Shehadah, Eyad,Jawhar, Mohammad,Arabi, Mohammad Imad Eddin The Korean Society of Plant Pathology 2014 Plant Pathology Journal Vol.30 No.4

Leaf scald caused by the infection of Rhynchosporium secalis, is a worldwide crop disease resulting in significant loss of barley yield. In this study, a systematic sequencing of expressed sequence tags (ESTs) was chosen to obtain a global picture of the assembly of genes involved in pathogenesis. To identify a large number of plant ESTs, which are induced at different time points, an amplified fragment length polymorphism (AFLP) display of complementary DNA (cDNA) was utilized. Transcriptional changes of 140 ESTs were observed, of which 19 have no previously described function. Functional annotation of the transcripts revealed a variety of infection-induced host genes encoding classical pathogenesis-related (PR) or genes that play a role in the signal transduction pathway. The expression analyses by a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) revealed that Rar1 and Rpg4 are defense inducible genes, and were consistent with the cDNA-AFLP data in their expression patterns. Hence, the here presented transcriptomic approach provides novel global catalogue of genes not currently represented in the EST databases.

Transcriptome Analysis of the Barley-Rhynchosporium secalis Interaction

Antonious Al-Daoude,Amina Shoaib,Eyad Al-Shehadah,Mohammad Jawhar,Mohammad Imad Eddin Arabi 한국식물병리학회 2014 Plant Pathology Journal Vol.30 No.4

Leaf scald caused by the infection of Rhynchosporium secalis, is a worldwide crop disease resulting in significant loss of barley yield. In this study, a systematic sequencing of expressed sequence tags (ESTs) was chosen to obtain a global picture of the assembly of genes involved in pathogenesis. To identify a large number of plant ESTs, which are induced at different time points, an amplified fragment length polymorphism (AFLP) display of complementary DNA (cDNA) was utilized. Transcriptional changes of 140 ESTs were observed, of which 19 have no previously described function. Functional annotation of the transcripts revealed a variety of infection-induced host genes encoding classical pathogenesis- related (PR) or genes that play a role in the signal transduction pathway. The expression analyses by a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) revealed that Rar1 and Rpg4 are defense inducible genes, and were consistent with the cDNA-AFLP data in their expression patterns. Hence, the here presented transcriptomic approach provides novel global catalogue of genes not currently represented in the EST databases.

Cloning, Expression and Hormonal Regulation of Steroidogenic Acute Regulatory Protein Gene in Buffalo Ovary

Malhotra, Nupur,Singh, Dheer,Sharma, M.K. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.2

In mammalian ovary, steroidogenic acute regulatory (StAR) protein mediates the true rate-limiting step of transport of cholesterol from outer to inner mitochondrial membrane. Appropriate expression of StAR gene represents an indispensable component of steroidogenesis and its regulation has been found to be species specific. However, limited information is available regarding StAR gene expression during estrous cycle in buffalo ovary. In the present study, expression, localization and hormonal regulation of StAR mRNA were analyzed by semi-quantitative RT-PCR in buffalo ovary and partial cDNA was cloned. Total RNA was isolated from whole follicles of different sizes, granulosa cells from different size follicles and postovulatory structures like corpus luteum and Corpus albicans. Semi-quantitative RT-PCR analyses showed StAR mRNA expression in the postovulatory structure, corpus luteum. No StAR mRNA was detected in total RNA isolated from whole follicles of different size including the preovulatory follicle (>9 mm in diameter). However, granulosa cells isolated from preovulatory follicles showed the moderate expression of StAR mRNA. To assess the hormonal regulation of StAR mRNA, primary culture of buffalo granulosa cells were treated with FSH (100 ng/ml) alone or along with IGF-I (100 ng/ml) for 12 to 18 h. The abundance of StAR mRNA increased in cells treated with FSH alone or FSH with IGF-I. However, effect of FSH with IGF-I on mRNA expression was found highly significant (p<0.01). In conclusion, differential expression of StAR messages was observed during estrous cycle in buffalo ovary. Also, there was a synergistic action of IGF-I on FSH stimulation of StAR gene.

Antioxidant and Anti-proliferative Activities of Different Solvent Fractions from Methanol Extracts of Sparganiun stoloniferum

Ming-Lu Xu,Lan Wang,Gui-Fang Xu,Jian-He Hu,Myeong-Hyeon Wang 한국원예학회 2010 Horticulture, Environment, and Biotechnology Vol.51 No.6

The antioxidant and antiproliferation effects of fractions from methanol extract of Sparganiun stoloniferum were investigated in this study. The ethyl acetate fraction of the methanol extract showed significant antioxidant activity against α,?α-diphenyl-picrylhydrazyl (DPPH) and hydroxyl ( · OH) radical scavenging, and it showed greater reducing power than the n-hexane, dichloromethane, n-butanol and water fractions. Qualitative HPLC analysis was performed on the extractions. The high content of phenolics in ethyl acetate fractions were believed to be responsible for its antioxidant activity, which was the strongest in all five fractions. Moreover, the ethyl acetate fraction showed a dose-dependent inhibition of cell proliferation with an EC?? of 139.21 ± 0.54 μg · mL?¹ against the human colon cell, HT-29. The anti-proliferative effects of the ethyl acetate fraction were associated with apoptosis. Semi-quantitative RT-PCR analysis indicates that, after treatment with the ethyl acetate fraction, the expression of caspase-3, p53, and Bax in HT-29 cells was significantly up-regulated and c-myc expression was induced. These findings demonstrate that the ethyl acetate fraction of S. stoloniferum has potent antioxidant and antiproliferation effects and induces apoptosis. Thus, the extract of S. stoloniferum may be considered as a potential functional food resource and complementary medicinal supplement.

cDNA-AFLP analysis reveals differential gene expression in response to salt stress in Brachypodium distachyon

김대연,홍민정,장지희,서용원 한국유전학회 2012 Genes & Genomics Vol.34 No.5

Environmental stresses such as drought, salinity, cold, and heat negatively affect the growth of plants and productivity of crops. The mechanism of salt tolerance is one of the most important fields in plant science, and our understanding of this process must be improved in order to increase agricultural crop production. In our study, we identified salt stress-responsive transcripts using the cDNA-AFLP technique. The obtained transcripts were further analyzed by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) under various abiotic stresses and hormone treatments. Among 87 transcript-derived fragments (TDFs) that were classified based on their presence or absence (qualitative variants) or differential expression (quantitative variants), we identified 32 TDFs that corresponded to Brachypodium genes with locus name. These clones are involved in various molecular functions and have transferase, protein binding, nucleotide binding, transporter,protein kinase, catalytic, hydrolase, RNA binding, and enzyme regulation activities. Further, the expression patterns of up-regulated 9 salt stress-response genes in cDNA-AFLP experiments were evaluated through semi-quantitative RT-PCR. Those genes were involved in signaling cascades [the casein kinase I (CKI)-like protein], regulation of enzyme activity [protein phosphatase 2C (PP2C) gene], phospholipid asymmetry [aminophospholipid ATPase (ALA)], cellular ion homeostasis [calcium-transporting ATPase, potassium transporter,calcium-binding protein (CBP)], and plant growth and development [pentatricopeptide repeat-containing protein (PPRP),2-oxoglutarate (2OG) and Fe(II)-dependent oxygenases] and the putative roles of the identified TDFs involved in salt stress mechanisms are discussed. A better understanding of the mechanisms of salt stress tolerance and salt stress response genes in Brachypodium would be very useful for the breeding and genetic engineering of salt tolerance varieties in other Poaceae families, including wheat, barley, and rice.

Polygalacturonase (PG) Gene Expression in Musa acuminata Cultivars from Kerala

Thulasy Gayathri,Ashalatha Sunkarankutty Nair,Vidhu Aniruddha Sane 한국식품과학회 2013 Food Science and Biotechnology Vol.22 No.6

Banana cannot be preserved for a long timeafter harvesting due to a short shelf life. Fruit softening isassociated with textural changes due to disassembly of theprimary cell wall and modification of the structure andcomposition of various polysaccharides. Cell wall degradationis caused by the action of various cell wall hydrolaseenzymes. Polygalacturonase (PG) is the key enzymeinvolved in this process. The ripening period is different incultivars maintained under domesticated cultivation inKerala. PG activity was profiled in eight Musa acuminatacultivars from Kerala and expression analysis of the PGgene was accomplished using semi-quantitative RT-PCR. Maximum PG activity was observed in Palayankodan andminimum activity was observed in Kadali. Gene expressionanalysis showed variation between ethylene treated fruitsand controls in Palayankodan, whereas the expressionpatterns in Kadali were similar. The fruit softening processis cultivar specific.

Antioxidant and Nitric Oxide Release Inhibition Activities of Methanolic Extract from Clerodendrum cyrtophyllum Turcz

Huan Liu,Lan Wang,왕명현 한국원예학회 2011 Horticulture, Environment, and Biotechnology Vol.52 No.3

Antioxidant and anti-inflammatory activities of methanolic extract of Clerodendrum cyrtophyllum Turcz (ECT) were investigated in the present study. ECT showed high phenolic content (223 μg・GAE-1 equivalent) and strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (IC50 = 52.74 μg・mL-1) and reduced power ability. Furthermore, ECT exhibited significant protective ability in H2O2/Fe3+/ascorbic acid-induced DNA and protein damage. Moreover, ECT inhibited NO production by 47.15% at 0.8 mg・mL-1 in lipopolysaccharides-stimulated RAW264.7 macrophages. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis showed that the expression of iNOS and COX-2 in RAW264.7 cells was down-regulated after treatment with increasing concentration of ECT. In conclusion, ECT has shown potent antioxidant and anti-inflammatory activities could be considered as a source of health promoting antioxidant and complementary medicinal supplement.

Oleuropein Induces Apoptosis Via the p53 Pathway in Breast Cancer Cells

Hassan, Zeinab Korany,Elamin, Maha Hussein,Omer, Sawsan Ali,Daghestani, Maha Hassan,Al-Olayan, Ebtesam Salah,Elobeid, Mai AbdelRahman,Virk, Promy Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.11

Background: Breast cancer is a major health problem worldwide. Olive oil induces apoptosis in some cancer cells due to phenolic compounds like oleuropein. Although oleuropein has anticancer activity, the underlying mechanisms of action remain unknown. The study aimed to assess the mechanism of oleuropin-induced breast cancer cell apoptosis. Materials and Methods: p53, Bcl-2 and Bax gene expression was evaluated by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) in luminal MCF-7 cells. Results: Oleuropein-induced apoptosis was accompanied by up-regulation of both p53 and Bax gene expression levels and down-regulation in Bcl2. Conclusions: Oleuropein induces apoptosis in breast tumour cells via a p53-dependent pathway mediated by Bax and Bcl2 genes. Therefore, oleuropein may have therapeutic potential in breast cancer patients by inducing apoptosis via activation of the p53 pathway.