Proteolytic machinery involved in MICA and MICB processing in cervical Cancer
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1
CUCS, Universidad de Guadalajara, Departamento de Fisiología, Mexico
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2
CUCS, Universidad de Guadalajara, Departamento de Biología Molecular y Genómica, Mexico
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3
CUCS, Universidad de Guadalajara, Departamento de Microbiología y Patología, Mexico
Introduction: MICA and MICB are upregulated in epithelial tumors. The activating NKG2D receptor, expressed on NK and cytotoxic T lymphocytes, recognizes MIC molecules leading to anti-tumor responses. However, it has been demonstrated that tumor cells use proteolytic cleavage to shed MICA/B as a mechanism to evade immunosurveillance. Objective: To identify the expression profiles of ADAM10, ADAM17, MMP9 and Erp5 in cervical cancer cell lines and to associate these with soluble MICA/B levels. Methods: Three different cervical cancer cell lines were used (HeLa, SiHa and C-33A) as well as non-tumorigenic keratinocytes (HaCaT). The transcriptional expression of ADAM10, ADAM17, MMP9 and Erp5 was evaluated using qRT-PCR. Identification at the protein level was assessed by Western blot. MMP9 activity was detected through zymography. Soluble MICA/B were quantified by ELISA. Results: All cell lines, including HaCaT, express the proteolytic machinery capable of shedding MICA/B. Interestingly, it was observed that the ADAM10 active form, and its zymogen, are mainly expressed in HPV-positive HeLa and SiHa cells, as compared to C33A, an HPV-negative tumorigenic cell line. The strongest MMP9 enzymatic activity was found in SiHa cells, which interestingly showed the highest soluble MICA/B levels. HaCaT did not show significant soluble MICA/B. Conclusions: While proteolytic machinery was present in all cell lines, soluble MICA/B was only found in tumor cell supernatants. This finding suggests that MICA/B processing is more complicated than expected and that additional mechanisms (such as different MICA/B variants, glycosylation or palmitoylation states) could facilitate the proteolytic cleavage from the tumor cell surface.
References
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Keywords:
NK cells,
NKG2D receptor,
NKG2D ligands,
shedding,
Metalloproteases,
cervical cancer
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Immune-mediated disease pathogenesis
Citation:
Garcia Chagollan
M,
Bueno-Topete
MR,
Fujimura-Hernandez
S,
Fafutis-Morris
M,
Franco-Topete
RA,
Bastidas-Ramírez
BE and
Del Toro-Arreola
S
(2013). Proteolytic machinery involved in MICA and MICB processing in cervical Cancer.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00308
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Received:
18 Mar 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Dr. Susana Del Toro-Arreola, CUCS, Universidad de Guadalajara, Departamento de Fisiología, Guadalajara, Mexico, susanadeltoro@hotmail.com