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Article: A protein-based set of reference markers for liver tissues and hepatocellular carcinoma

TitleA protein-based set of reference markers for liver tissues and hepatocellular carcinoma
Authors
Issue Date2009
PublisherBioMed Central Ltd. The Journal's web site is located at http://www.biomedcentral.com/bmccancer/
Citation
Bmc Cancer, 2009, v. 9, p. 309 How to Cite?
AbstractBackground: During the last decade, investigations have focused on revealing genes or proteins that are involved in HCC carcinogenesis using either genetic or proteomic techniques. However, these studies are overshadowed by a lack of good internal reference standards. The need to identify "housekeeping" markers, whose expression is stable in various experimental and clinical conditions, is therefore of the utmost clinical relevance in quantitative studies. This is the first study employed 2-DE analysis to screen for potential reference markers and aims to correlate the abundance of these proteins with their level of transcript expression. Methods: A Chinese cohort of 224 liver tissues samples (105 cancerous, 103 non-tumourous cirrhotic, and 16 normal) was profiled using 2-DE analysis. Expression of the potential reference markers was confirmed by western blot, immunohistochemistry and real-time quantitative PCR. geNorm algorithm was employed for gene stability measure of the identified reference markers. Results: The expression levels of three protein markers beta-actin (ACTB), heat shock protein 60 (HSP60), and protein disulphide isomerase (PDI) were found to be stable using p-values (p > 0.99) as a ranking tool in all 224 human liver tissues examined by 2-DE analysis. Of high importance, ACTB and HSP 60 were successfully validated at both protein and mRNA levels in human hepatic tissues by western blot, immunohistochemistry and real-time quantitative PCR. In addition, no significant correlation of these markers with any clinicopathological features of HCC and cirrhosis was found. Gene stability measure of these two markers with other conventionally applied housekeeping genes was assessed by the geNorm algorithm, which ranked ACTB and HSP60 as the most stable genes among this cohort of clinical samples. Conclusion: Our findings identified 2 reference markers that exhibited stable expression across human liver tissues with different conditions thus should be regarded as reliable reference moieties for normalisation of gene and protein expression in clinical research employing human hepatic tissues. © 2009 Sun et al; licensee BioMed Central Ltd.
Persistent Identifierhttp://hdl.handle.net/10722/83212
ISSN
2015 Impact Factor: 3.265
2015 SCImago Journal Rankings: 1.627
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
Innovation and Technology CommissionITS/120/07
University Research Committee of the University of Hong Kong
Sun Chieh Yeh Research Foundation for Hepatobiliary and Pancreatic Surgery
Funding Information:

The authors gratefully acknowledge the clinical support kindly offered by Professor S. T. Fan and clinical assistance provided by Ashley Wong, Department of Surgery, The University of Hong Kong. Fiona Salway of The Manchester Interdisciplinary Biocentre, UK is thanked for technical assistance, discussions and input. The work was supported by grants from the Innovation and Technology Commission (ITS/120/07) and the University Research Committee of the University of Hong Kong and Sun Chieh Yeh Research Foundation for Hepatobiliary and Pancreatic Surgery.

References
Grants

 

DC FieldValueLanguage
dc.contributor.authorSun, Sen_HK
dc.contributor.authorYi, Xen_HK
dc.contributor.authorPoon, RTPen_HK
dc.contributor.authorYeung, Cen_HK
dc.contributor.authorDay, PJRen_HK
dc.contributor.authorLuk, JMen_HK
dc.date.accessioned2010-09-06T08:38:20Z-
dc.date.available2010-09-06T08:38:20Z-
dc.date.issued2009en_HK
dc.identifier.citationBmc Cancer, 2009, v. 9, p. 309en_HK
dc.identifier.issn1471-2407en_HK
dc.identifier.urihttp://hdl.handle.net/10722/83212-
dc.description.abstractBackground: During the last decade, investigations have focused on revealing genes or proteins that are involved in HCC carcinogenesis using either genetic or proteomic techniques. However, these studies are overshadowed by a lack of good internal reference standards. The need to identify "housekeeping" markers, whose expression is stable in various experimental and clinical conditions, is therefore of the utmost clinical relevance in quantitative studies. This is the first study employed 2-DE analysis to screen for potential reference markers and aims to correlate the abundance of these proteins with their level of transcript expression. Methods: A Chinese cohort of 224 liver tissues samples (105 cancerous, 103 non-tumourous cirrhotic, and 16 normal) was profiled using 2-DE analysis. Expression of the potential reference markers was confirmed by western blot, immunohistochemistry and real-time quantitative PCR. geNorm algorithm was employed for gene stability measure of the identified reference markers. Results: The expression levels of three protein markers beta-actin (ACTB), heat shock protein 60 (HSP60), and protein disulphide isomerase (PDI) were found to be stable using p-values (p > 0.99) as a ranking tool in all 224 human liver tissues examined by 2-DE analysis. Of high importance, ACTB and HSP 60 were successfully validated at both protein and mRNA levels in human hepatic tissues by western blot, immunohistochemistry and real-time quantitative PCR. In addition, no significant correlation of these markers with any clinicopathological features of HCC and cirrhosis was found. Gene stability measure of these two markers with other conventionally applied housekeeping genes was assessed by the geNorm algorithm, which ranked ACTB and HSP60 as the most stable genes among this cohort of clinical samples. Conclusion: Our findings identified 2 reference markers that exhibited stable expression across human liver tissues with different conditions thus should be regarded as reliable reference moieties for normalisation of gene and protein expression in clinical research employing human hepatic tissues. © 2009 Sun et al; licensee BioMed Central Ltd.en_HK
dc.languageengen_HK
dc.publisherBioMed Central Ltd. The Journal's web site is located at http://www.biomedcentral.com/bmccancer/en_HK
dc.relation.ispartofBMC Canceren_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshActins - genetics - metabolism-
dc.subject.meshCarcinoma, Hepatocellular - genetics - metabolism-
dc.subject.meshChaperonin 60 - genetics - metabolism-
dc.subject.meshElectrophoresis, Gel, Two-Dimensional - methods - standards-
dc.subject.meshLiver Neoplasms - genetics - metabolism-
dc.titleA protein-based set of reference markers for liver tissues and hepatocellular carcinomaen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1471-2407&volume=9&spage=309&epage=&date=2009&atitle=A+protein-based+set+of+reference+markers+for+liver+tissues+and+hepatocellular+carcinomaen_HK
dc.identifier.emailPoon, RTP: poontp@hkucc.hku.hken_HK
dc.identifier.emailLuk, JM: jmluk@hkucc.hku.hken_HK
dc.identifier.authorityPoon, RTP=rp00446en_HK
dc.identifier.authorityLuk, JM=rp00349en_HK
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1186/1471-2407-9-309en_HK
dc.identifier.pmid19725976-
dc.identifier.pmcidPMC2742551-
dc.identifier.scopuseid_2-s2.0-70349854558en_HK
dc.identifier.hkuros167141en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-70349854558&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume9en_HK
dc.identifier.spage309en_HK
dc.identifier.epage309en_HK
dc.identifier.isiWOS:000270309200002-
dc.publisher.placeUnited Kingdomen_HK
dc.relation.projectLiver cancer biomarker test: diagnostic use of CDH17 monoclonal antibodies-
dc.identifier.scopusauthoridSun, S=21740136100en_HK
dc.identifier.scopusauthoridYi, X=55138104500en_HK
dc.identifier.scopusauthoridPoon, RTP=7103097223en_HK
dc.identifier.scopusauthoridYeung, C=26531966700en_HK
dc.identifier.scopusauthoridDay, PJR=7202148832en_HK
dc.identifier.scopusauthoridLuk, JM=7006777791en_HK
dc.identifier.citeulike5719526-

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