NDLI: Immunohistochemical localization of hTERT protein in human tissues

Content Provider	Springer Nature Link
Author	Yan, Pu Benhattar, Jean Seelentag, Walter Stehle, Jean Christophe Bosman, Fred T.
Copyright Year	2004
Abstract	Telomerase is a ribonucleoprotein complex mainly composed of a reverse transcriptase catalytic subunit (telomerase reverse transcriptase gene, hTERT) that copies a template region of its RNA subunit to the end of the telomere. For detecting telomerase activity in a tissue specimen the TRAP assay is a relatively sensitive and specific method, but it can be used only on fresh tissue extracts and offers no information at the single cell level. Immunohistochemistry (IHC) allows to detect hTERT protein expression at an individual cell level in human tissues. We have tested commercially available anti-hTERT antibodies in formalin-fixed and paraffin-embedded human tissues by IHC. Only one monoclonal antibody (NCL-hTERT; Novacastra) was sufficiently specific and this was applied to human tissues in which telomerase activity had been shown by TRAP assay and hTERT mRNA expression by RT-PCR. hTERT protein localized diffusely in the nucleoplasm and more intensely in the nucleoli of cancer cells and proliferating normal cells. Mitotic cells showed diffuse staining of the entire cell. Granular cytoplasmic staining was occasionally found in some tumor cells. In telomerase-positive tumors not all the tumor cells showed hTERT immunoreactivity. A significantly heterogeneous hTERT protein expression was observed in human tumor tissues. The hTERT immunostaining in fixed tissues was concordant with telomerase activity and hTERT mRNA expression in corresponding non-fixed samples. Quantitative RT-PCR of microdissected sections showed that hTERT mRNA expression was higher in cells with nuclear expression than in those with cytoplasmic expression. Double staining with the M30 antibody showed that a subpopulation of hTERT-negative cells is apoptotic. We conclude that: (1) hTERT protein can be detected by IHC in fixed human tissues, but the choice of the antibody, tissue processing, and reaction conditions are critical, (2) hTERT protein localizes in the nucleoplasm, more strongly in the nucleolus, and occasionally in the cytoplasm, (3) telomerase-positive tumors show significant heterogeneity of hTERT protein expression, and (4) a subpopulation of hTERT protein negative tumor cells is identified as apoptotic cells.
Starting Page	391
Ending Page	397
Page Count	7
File Format	PDF
ISSN	09486143
Journal	Histochemistry and Cell Biology
Volume Number	121
Issue Number	5
e-ISSN	1432119X
Language	English
Publisher	Springer-Verlag
Publisher Date	2004-05-07
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	hTERT Immunohistochemistry Telomerase TRAP assay RT-PCR Apoptotic cells
Content Type	Text
Resource Type	Article
Subject	Cell Biology Histology Molecular Biology Medical Laboratory Technology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Protein phosphatase 2Cα expression in normal human tissues: an immunohistochemical study

Immunohistochemical Detection of Telomerase (hTERT) Protein in Human Cancer Tissues and a Subset of Cells in Normal Tissues

Immunohistochemical localization of heparan sulfate proteoglycan in human gastrointestinal tract

Novel distribution of adrenomedullin-immunoreactive cells in human tissues

Immunohistochemical localization of sodium–potassium ATPase in human normal stomach and gastric adenocarcinomas

Localization of human airway trypsin-like protease in the airway: an immunohistochemical study

Differential expression of annexins I, II and IV in human tissues: an immunohistochemical study

The normal cellular prion protein (PrPc) is strongly expressed in bovine endocrine pancreas

Juxtamembrane localization of the protein phosphatase-1 inhibitor protein PHI-1 in smooth muscle cells

Immunohistochemical localization of hTERT protein in human tissues

Similar Documents

Protein phosphatase 2Cα expression in normal human tissues: an immunohistochemical study

Immunohistochemical Detection of Telomerase (hTERT) Protein in Human Cancer Tissues and a Subset of Cells in Normal Tissues

Immunohistochemical localization of heparan sulfate proteoglycan in human gastrointestinal tract

Novel distribution of adrenomedullin-immunoreactive cells in human tissues

Immunohistochemical localization of sodium–potassium ATPase in human normal stomach and gastric adenocarcinomas

Localization of human airway trypsin-like protease in the airway: an immunohistochemical study

Differential expression of annexins I, II and IV in human tissues: an immunohistochemical study

The normal cellular prion protein (PrPc) is strongly expressed in bovine endocrine pancreas

Juxtamembrane localization of the protein phosphatase-1 inhibitor protein PHI-1 in smooth muscle cells

Immunohistochemical localization of hTERT protein in human tissues