Gene Expression 15(5-6) Abstracts

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Gene Expression, Vol. 15, pp. 199–206, 2013
1052-2166/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/105221613X13571653093123
E-ISSN 1555-3884
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

RNA Interference-Mediated Knockdown of DGAT1 Decreases Triglyceride Content of Bovine Mammary Epithelial Cell Line

Chunyan Lu,1 Runjun Yang,1 Binglei Shen, Hassan Osman, Yonghong Zhang, Shouqing Yan, Liying Zhang, and Zhihui Zhao

College of Animal Science, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, Jilin Province, China

Diacylglyceroltransferase-1 (DGAT1) expresses in nearly all tissues, including the mammary gland. Mice lacking DGAT1 exhibit decreased triglyceride content in mammary tissue, and are resistant to diet-induced obesity and diabetes mellitus. Thus, DGAT1 has received considerable attention. In the present study, the function of DGAT1 was examined by liposome mediated RNA interference (RNAi) to knockdown the expression of endogenous DGAT1 expression in bovine mammary epithelial cells (BMEC) and the changes of the biological functions of cells were analyzed. The mRNA and protein levels, intracellular triglyceride (TG) content, and total protein of BMECs were analyzed by real-time PCR, Western blot, TG kit, and ultraviolet spectrophotometer, respectively, before and after RNAi treatment. The results indicated that knockdown of DGAT1 expression significantly reduced TG content in BMECs. This study further confirmed the importance of DGAT1 in triglyceride synthesis in bovine mammary tissue.

Key words: DGAT1; RNA interference (RNAi); Triglyceride (TG); Mammary epithelial cell

1These authors provided equal contribution to this work.
Address correspondence to Zhihui Zhao, College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, 5333 Xi’an Road, Changchun, Jilin Province, 130062, China. Tel: +86-431-87836156; Fax: +86-431-87836156; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Liying Zhang, College of Animal Science and Veterinary Medicine, and Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, 5333 Xi’an Road, Changchun, Jilin Province, 130062, China. Tel: +86-431-87836156; Fax: +86-431-87836156; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Gene Expression, Vol. 15, pp. 207–214, 2013
1052-2166/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/105221613X13571653093169
E-ISSN 1555-3884
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Involvement of ITIH5, a Candidate Gene for Congenital Uterovaginal Aplasia (Mayer-Rokitansky-Küster-Hauser Syndrome), in Female Genital Tract Development

Karine Morcel,*† Tanguy Watrin,* Frédérique Jaffre,*† Stéphane Deschamps,*‡ Francis Omilli,*‡ Isabelle Pellerin,*‡ Jean Levêque,†‡ and Daniel Guerrier*‡

*
CNRS, UMR 6290-IGDR, Rennes, France
†CHU Rennes, Pôle d’Obstétrique Gynécologie et Médecine de la Reproduction, Rennes, France
‡University of Rennes1, UEB, UMS 3480-Biosit, Faculté de Médecine, Rennes, France

The ITI (inter-trypsine inhibitor) gene family includes five genes (ITIH1 to ITIH5) that encode proteins involved in the dynamics of the extracellular matrix (ECM). ITIH5 was found inactivated by partial deletion in a case of congenital uterovaginal aplasia, a human rare disease also called Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome. The aim of the present study was to analyze the expression of ITIH5 in the uterus in adult life and during embryogenesis in order to establish the involvement of this gene in both normal and pathological conditions of uterus development. This was achieved in mice by reverse transcription-quantitative PCR, whole-mount hybridization, and Western blot analysis. Itih5 expression was much stronger in female genital tract primordial (Müllerian ducts) and derivatives than elsewhere in the body. This gene was strongly expressed during pregnancy and development of the female genital tract, indicating that the encoded protein probably had an important function in the uterus during these periods. Two different specific isoforms of the protein were detected in Müllerian derivatives during embryogenesis and in adults. Although ITIH genes are expected to be predominantly expressed in the liver, ITIH5 is mainly expressed in the uterus during development and adult life. This tends to indicate an additional and specific role of this gene in the female reproductive tract, and furthermore reinforces ITIH5 as a putative candidate gene for MRKH syndrome.

Key words: ITIH5; Extracellular matrix; Female; Mouse; Congenital uterovaginal aplasia; MRKH syndrome

Address correspondence to Dr. Karine Morcel, Pôle d’Obstétrique Gynécologie et Médecine de la Reproduction, CHU Hôpital Sud, 16 boulevard de Bulgarie BP 90437, 35203 Rennes Cedex 2, France. Tel: +33 (0)2 99 26 67 09; Fax: +33 (0)2 99 26 67 35; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Gene Expression, Vol. 15, pp. 215–223, 2013
1052-2166/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/105221613X13571653093204
E-ISSN 1555-3884
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Identification of Genotypes of Plasmid-Encoded AmpC β-Lactamases From Clinical Isolates and Characterization of Mutations in Their Promoter and Attenuator Regions

Gui-Ling Li,* Li-Bo Duo,* Ying Luan,* Cheng-Ying Wang,† Wei-Ping Wang,† He-Guang Zhang,* Qi Sun,* and Gui-Yun Qi*

*Department of Medicine Laboratory, Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China
†Medicine Laboratory, Department of Urology Surgery, DaQing Oilfield General Hospital, DaQing, Heilongjiang, China

We investigated the occurrence of AmpC β-lactamases among Escherichia coli and Klebsiella pneumonia isolates and determined the genotype of plasmid-mediated AmpC β-lactamases at a medical center. The AmpC β-lactamase promoter and attenuator were amplified from chromosomal DNA of high AmpC-producing E. coli isolates and sequenced. Antibiotic screening and 3D extract tests showed the presence of AmpC β-lactamase in 3.56% of K. pneumoniae and 1.88% of E. coli isolates. Ten isolates (six K. pneumoniae and four E. coli) were positive for extended spectrum β-lactamase (ESBL) as indicated by the double disc diffusion method. DHA-1 plasmid-encoded AmpC β-lactamase was present in 10 K. pneumoniae isolates and four E.coli isolates. E. coli chromosomal AmpC β-lactamase carried polymorphisms in the −42, −32, and −18 bases of the promoter and in the +26 and +27 bases of the attenuator, which may play a role in antibiotic resistance. The observed mutations may have clinical implications for the management of antibiotic-resistant infections.

Key words: Klebsiella pneumoniae; Escherichia coli; AmpC β-lactamases; DHA-1; Inducible

Address correspondence to Li-Bo Duo, Department of Medicine Laboratory, Second Affiliated Hospital of Harbin Medical University, 148 Baojian Road, Harbin, Heilongjiang, 150086, China. Tel: +86-0451-86605363; Fax: +86-0451-86296362; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Gene Expression, Vol. 15, pp. 225–234, 2013
1052-2166/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/105221613X13571653093240
E-ISSN 1555-3884
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Statins Cause Profound Effects on Gene Expression in Human Cancer Cells In Vitro: The Role of Membrane Microdomains

David John Garnett and Trevor James Greenhough

Structural Biology Research Group, Institute of Science Technology in Medicine, Keele University, Keele, Staffordshire, UK

There is increasing evidence that statin treatment can be beneficial in certain cancer patients. To determine if these benefits are a direct result of the cholesterol-lowering effects of statins or a result of secondary, protein transcription effects, the impacts of pravastatin and a cholesterol sequestrating agent methyl-β-cyclodextrin (MβCD) on mRNA expression in the breast cancer cell MDA-MB-231 and the lung carcinoma cell Calu-1 have been compared by microarray techniques. The effects of these agents on cholesterol-rich rafts and caveolae, which have significance in cancer signaling, have also been examined. Both treatments caused a general downregulation of not only signal transduction including cancer pathway proteins, but also apoptosis and chemokine pathways, with statins impacting 35 genes by twofold or greater in MDA-MB-231 and >300 genes in Calu-1. These manifold dysregulations could also explain the various side effects reportedly caused by statins. MβCD produced far fewer statistical events than pravastatin in the breast cancer line but many more in the lung cell line. Pravastatin increased expression of CAV1 but caveolae density decreased and overall raft density was unaffected. MβCD also caused an increase in CAV1 expression and reduced the prevalence of both rafts and caveolae. It is proposed that sequestration of cholesterol from the membrane by MβCD is not equivalent to blockade of the cholesterol pathway and causes different effects on microdomain-mediated signal transduction dependant on the cell line. The profound effects of statins on mRNA expression can be explained by the failure of caveolin-1 to properly complex with cholesterol in an altered sterol environment, with caveolae acting as the main loci for signaling directed towards those transcription processes unaffected by MβCD. Targeted inhibition of the postmevalonate pathway could offer an opportunity to specifically reduce caveolae-based signaling in cancer cells. The observed impact of pravastatin on gene expression may explain the pleiotropic effects of statins when they are used as adjuvants in chemotherapy and suggests impact on gene expression as a possible cause of side effects from statin use.

Key words: Cancer; Gene expression; Pravastatin; Rafts; Caveolae

Address correspondence to David John Garnett, Structural Biology Research Group, Institute of Science Technology in Medicine, Keele University, Keele, Staffordshire, ST5 5BG UK. Tel: 01782 554718; Fax: 01782 734637; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Gene Expression, Vol. 15, pp. 235–241, 2013
1052-2166/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/105221613X13571653093286
E-ISSN 1555-3884
Copyright © 2013 Cognizant Comm. Corp.
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Cystathionine Gamma-Lyase Expression Is Regulated by Exogenous Hydrogen Peroxide in the Mammalian Cells

Maoxian Wang,*† Zhanyun Guo,* and Shilong Wang*

*College of Life sciences and Technology, Tongji University, Shanghai, P.R. China
†Department of Biology, Hanshan Normal University, Chaozhou, P.R. China

Hydrogen sulfide (H2S), as an endogenous signaling molecule in mammals, shows a variety of biological effects. Cystathionine gamma-lyase (CSE)/H2S pathway has been implicated in scavenging reactive oxygen species (ROS) in the mammalian cells. Therefore, we first investigated the regulatory effects of exogenously applied hydrogen peroxide (H2O2) on CSE expression in the mammalian cells. African green monkey kidney fibroblastlike cells (COS-7 cells) or human embryonic kidney 293 cells (HEK 293 cells) were transfected with CSE promoter-luciferase reporter constructs and treated with H2O2 of 1, 5, and 10 μM for 0.5 and 1.5 h at 37°C. The transfected cells were assayed for firefly luciferase activities normalized by Renilla luciferase activity. Human lung adenocarcinoma cells (A549 cells) or human liver cancer cells (SMMC-7721 cells) were treated with H2O2 of 1, 5, and 10 μM for 0.5 and 1.5 h at 37°C, and were then harvested and analyzed by Western blotting and quantitative RT-PCR. Our results showed that the treatment of a medium concentration (5 μM) of H2O2 at a longer time (1.5 h) upregulated CSE expression in the mammalian cells at the levels of the promoter, message RNA, and protein. Collectively, exogenously applied H2O2 can not only markedly affect CSE mRNA and protein expression, but also can affect the CSE promoter activity in the mammalian cells. Our observations indicate that that exogenous H2O2 can upregulate the expression of the CSE gene in the mammalian cells, which will provide the possibility of the scavenging effect of the CSE gene indirectly on ROS in the mammalian cells. However, the regulatory mechanism involved in the effects of exogenously applied H2O2 on CSE expression in the mammalian cells need be further studied.

Key words: Hydrogen sulfide; Cystathionine gamma-lyase; Hydrogen peroxide; Oxidative stress

Address correspondence to Shilong Wang, College of Life Sciences and Technology, Room 1119, Yixue Building, Tongji University, No 1239 Siping Road, Shanghai 200092, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Gene Expression, Vol. 15, pp. 243–253, 2013
1052-2166/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/105221613X13571653093321
E-ISSN 1555-3884
Copyright © 2013 Cognizant Comm. Corp.
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Computational Approaches for Identifying Cancer miRNA Expressions

Shubhra Sankar Ray,*† Jayanta Kumar Pal,* and Sankar K. Pal*†

*Center for Soft Computing Research, Indian Statistical Institute, Kolkata, India
†Machine Intelligence Unit, Indian Statistical Institute, Kolkata, India

MicroRNAs (miRNAs) play a major role in cancer development and also act as a key factor in many other diseases. In this investigation, we propose three methods for handling miRNA expressions. The first two methods determine whether a miRNA is indicating normal or cancer condition, and the third one determines how many miRNAs are supporting the cancer sample/patient. While Method 1 acts as a two-class classifier and is based on normalized average expression value, Method 2 also does the same and is based on the normalized average intraclass distance. Method 3 checks whether a miRNA belongs to the cancer class or not, provides the percentage of supporting miRNAs for a cancer patient, and is based on weighted normalized average intraclass distance. The values of the weights are determined using exhaustive search by maximizing the accuracy in training samples. The proposed methods are tested on the differentially regulated miRNAs in three types of cancers (breast, colon, and melanoma cancer). The performances of Method 1 and Method 2 are evaluated by F score, Matthews Correlation Coefficient (MCC), and plotting “1 − specificity versus sensitivity” in Receiver Operating Characteristic (ROC) space and are found to be superior to the kNN and SVM classifiers for breast, colon, and melanoma cancer data sets. It is also observed that both the sensitivity and the specificity of Method 1 and Method 2 are higher than 0.5. For the same data sets, Method 3 achieved an average accuracy of more than 98% in detecting the miRNAs, supporting the cancer condition.

Key words: Bioinformatics; MicroRNA expressions; Cancer; Pattern recognition

Address correspondence to Jayanta Kumar Pal, Junior Research Fellow, Center for Soft computing Research, Indian Statistical Institute, 203 B. T. Road, Kolkata-700108, India.Tel: +913325752048; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it