Document Detail


Bovine seminal ribonuclease is cytotoxic for both malignant and normal telomerase-positive cells.
MedLine Citation:
PMID:  16142325     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Bovine seminal-ribonuclease (BS-RNase) is a member of the 'ribonucleases with special biological actions' family since it possesses specific anti-tumour, anti-spermatogenic and embryotoxic activities and exerts an immunosuppressive effect on T lymphocytes. In previous studies it was demonstrated that BS-RNase induced apoptosis in proliferating, malignant and normal cells and that telomerase activity loss also caused apoptotic death in neoplastic cells. Since an obvious relationship between cell proliferation and telomerase activity exists, the aim of this work was to study if the pro-apoptotic cytotoxic action exerted by BS-RNase on proliferating malignant cells (HT29) and proliferating normal cells (PHA-stimulated lymphocytes) could be linked to a possible BS-RNase effect on telomerase activity. In BS-RNase-treated HT29 cells (Na-butyrate-differentiated or not) and human lymphocytes (proliferating or not), we investigated cell vitality (MTT method) and morphology (SEM), BS-RNase localization (immunofluorescence), telomerase activity (TRAP-ELISA method), hTR mRNA expression (RT-PCR), and hTERT levels (western blot). While no BS-RNase effect was detectable on not proliferating cells, a clear relationship was noticed between the diminished number of vital elements of both proliferating cell populations after treatment (48 h and 72 h for HT29 and PHA-stimulated lymphocytes, respectively) with 50 microg/ml BS-RNase and the decrease of their telomerase activity. At the same time, we found that hTR levels, the RNA subunit of telomerase, in proliferation-inhibited BS-RNase-treated cells were diminished. Moreover, by immunofluorescence technique, we detected BS-RNase in the HT29 cell nucleolus after 3-h treatment. Therefore, as hTR has been recently proven to co-fractionate with nucleoli, we hypothesize that a BS-RNase direct action on the telomerase hTR subunit could be a possible mechanism of action by which BS-RNase exerts its pro-apoptotic effects only on proliferating cells.
Authors:
Maria Viola; Massimo Libra; Daniela Callari; Fulvia Sinatra; Daniele Spada; Davide Noto; Giovanni Emmanuele; Fabrizio Romano; Maurizio Averna; Franca Maria Pezzino; Franca Stivala; Salvatore Travali
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  International journal of oncology     Volume:  27     ISSN:  1019-6439     ISO Abbreviation:  Int. J. Oncol.     Publication Date:  2005 Oct 
Date Detail:
Created Date:  2005-09-05     Completed Date:  2006-01-31     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9306042     Medline TA:  Int J Oncol     Country:  Greece    
Other Details:
Languages:  eng     Pagination:  1071-7     Citation Subset:  IM    
Affiliation:
Dipartimento di Chimica Biologica, Chimica Medica e Biologia Molecolare, Università degli Studi di Catania, Catania, Italy.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antineoplastic Agents / pharmacology
Apoptosis
Blotting, Western
Cattle
Cell Differentiation
Cell Line, Tumor
Cell Nucleolus / metabolism
Cell Proliferation
Down-Regulation
Endoribonucleases / chemistry*,  pharmacology*
Enzyme-Linked Immunosorbent Assay
Humans
Immunosuppressive Agents / pharmacology
Leukocytes, Mononuclear / cytology
Lymphocytes / metabolism
Microscopy, Electron, Scanning
Microscopy, Fluorescence
RNA / metabolism
RNA, Messenger / metabolism
Reverse Transcriptase Polymerase Chain Reaction
T-Lymphocytes / drug effects
Telomerase / biosynthesis*,  metabolism
Tetrazolium Salts / pharmacology
Thiazoles / pharmacology
Time Factors
Chemical
Reg. No./Substance:
0/Antineoplastic Agents; 0/Immunosuppressive Agents; 0/RNA, Messenger; 0/Tetrazolium Salts; 0/Thiazoles; 0/telomerase RNA; 298-93-1/thiazolyl blue; 63231-63-0/RNA; EC 2.7.7.49/Telomerase; EC 3.1.-/Endoribonucleases; EC 3.1.27.-/ribonuclease SPL

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