Immune Netw.  2004 Sep;4(3):184-189. 10.4110/in.2004.4.3.184.

TNF-alphaUp-regulated the Expression of HuR, a Prognostic Marker for Ovarian Cancer and Hu Syndrome, in BJAB Cells

• Lee KY

• KMID: 2167980
• DOI: http://doi.org/10.4110/in.2004.4.3.184

Abstract

BACKGROUND
Hu syndrome, a neurological disorder, is characterized by the remote effect of small cell lung cancer on the neural degeneration. The suspicious effectors for this disease are anti-Hu autoantibodies or Hu-related CD8+ T lymphocytes. Interestingly, the same effectors have been suggested to act against tumor growth and this phenomenon may represent natural tumor immunity. For these diagnostic and therapeutic reasons, the demand for antibodies against Hu protein is rapidly growing. METHODS: Polyclonal and monoclonal antibodies were generated using recombinant HuR protein. Western blot analyses were performed to check the specificity of generated antibodies using various recombinant proteins and cell lysates. Extracellular stimuli for HuR expression had been searched and HuR-associated proteins were isolated from polysome lysates and then separated in a 2-dimensional gel. RESULTS: Polyclonal and monoclonal antibodies against HuR protein were generated and these antibodies showed HuR specificity. Antibodies were also useful to detect and immunoprecipitate endogenous HuR protein in Jurkat and BJAB. This report also revealed that TNF-alphatreatment in BJAB up-regulated HuR expression. Lastly, protein profile in HuR-associated mRNA- protein complexes was mapped by 2-dimensional gel electrophoresis. CONCLUSION: This study reported that new antibodies against HuR protein were successfully generated. Currently, project to develop a diagnostic kit is in process. Also, this report showed that TNF-alphaup-regulated HuR expression in BJAB and protein profile associated with HuR protein was mapped.

Keyword

Hu syndrome; HuR; antibody; TNF-alpha; mRNP

MeSH Terms

Antibodies
Antibodies, Monoclonal
Autoantibodies
Blotting, Western
Electrophoresis
ELAV Proteins
ELAV-Like Protein 1
Nervous System Diseases
Ovarian Neoplasms*
Polyribosomes
Recombinant Proteins
Sensitivity and Specificity
Small Cell Lung Carcinoma
T-Lymphocytes
Tumor Necrosis Factor-alpha
Antibodies
Antibodies, Monoclonal
Autoantibodies
Recombinant Proteins
Tumor Necrosis Factor-alpha