Frequent co-expression of EGFR and NeuGcGM3 ganglioside in cancer: it’s potential therapeutic implications
Clin Exp Metastasis
Frequent co-expression of EGFR and NeuGcGM3 ganglioside in cancer: it's potential therapeutic implications
Addys Gonza´lez Palomo 0 1 2 3
Rance´s Blanco Santana 0 1 2 3
Xiomara Escobar Pe´rez 0 1 2 3
Damia´n Blanco Santana 0 1 2 3
Mariano Rolando Gabri 0 1 2 3
Kalet Leo´n Monzon 0 1 2 3
Adriana Carr Pe´rez 0 1 2 3
0 Department of Cell Biology and Tissues Banking, National Institute of Oncology and Radiology , 29 and F Street, Vedado, Plaza de la Revolucio ́n, P.O. Box 10400, Havana , Cuba
1 Center of Molecular Immunology (CIM) , 216 Street and 15 Avenue, Atabey, Playa, P.O. Box 16040, Habana , Cuba
2 & Addys Gonza ́lez Palomo
3 Laboratory of Molecular Oncology, Quilmes National University , R. Sa ́enz Pen ̃a 180, P.O. Box B1876BXD, Buenos Aires , Argentina
Interaction between epidermal growth factor receptor (EGFR) signaling with GM3 ganglioside expression has been previously described. However, little is known about EGFR and NeuGcGM3 co-expression in cancer patients and their therapeutic implications. In this paper, we evaluate the co-expression of EGFR and NeuGcGM3 ganglioside in tumors from 92 patients and in two spontaneous lung metastasis models of mice (Lewis lung carcinoma (3LL-D122) in C57BL/6 and mammary carcinoma (4T1) in BALB/c). As results, co-expression of EGFR and NeuGcGM3 ganglioside was frequently observed in 63 of 92 patients (68 %), independently of histological subtype. Moreover, EGFR is co-expressed with NeuGcGM3 ganglioside in the metastasis of 3LLD122 and 4T1 murine models. Such dual expression appears to be therapeutically relevant, since combined therapy with mAbs against these two molecules synergistically increase the survival of mice treated. Overall, our results suggest that NeuGcGM3 and EGFR may Addys Gonza´lez Palomo and Rance´s Blanco Santana have equally contributed to this work. Kalet Leo´n Monzon and Adriana Carr Pe´rez co-supervised this work.
EGFR; NeuGcGM3 Co-expression Pulmonary metastasis Combination therapy
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Most epithelial tumors overexpress the EGFR and their
activation is related with cancer progression. However,
tumors exhibit a diverse response to anti-EGFR therapies,
with resistance as common result of the treatment [1].
The N-Acetylneuramic acid (NeuAcGM3) ganglioside,
but not the N-glycolylneuramic acid (NeuGcGM3), is
usually detected in normal human tissues. However, many
human tumors express NeuGcGM3 ganglioside [2–7]. The
expression of NeuGcGM3 have been associated with a
worse prognosis in colon [8] and lung cancer [7, 9].
Differential association between EGFR signaling pathway and
GM3 ganglioside expression has been reported [10–13].
Overexpression of GM3 increase the proliferation of
carcinoma cells (A431) by ERK-independent signaling, in the
presence of urokinase plasminogen activator (uPA) and
their receptor (uPAr) [14]. Conversely, GM3 depletion
increase the EGFR phosphorylation and the
ERK-dependent cell proliferation becomes prevalent [14]. These
results provide a rational for a combined treatment
targeting simultaneously both EGFR and GM3 mediated
signaling pathways.
The Center of Molecular Immunology (CIM, Havana,
Cuba) have developed several immunotherapeutic projects
targeting separately both EGFR [15, 16] and NeuGcGM3
[17, 18]. Therefore, we evaluate the frequency of
coexpression of EGFR and NeuGcGM3 ganglioside in human
tumors and in two spontaneous lung metastasis models of
mice (Lewis lung carcinoma (3LL-D122) in C57BL/6 and
mammary carcinoma (4T1) in BALB/c). Moreover, we
perform an initial evaluation of the therapeutic implications
of targeting simultaneously both molecules, in lung
models.
Materials and methods
Patients samples
Sections of formalin-fixed and paraffin-embedded tumor
tissues from 92 patients were taken from the pathology
department of the National Institute of Oncology and
Radiobiology and Dr. ¨Manuel Fajardo¨ General Teaching
Hospital after receiving approved consent by the Ethical
Committee of the institute.
Lewis lung carcinoma (3LL-D122); mouse breast
adenocarcinoma cells (4T1); human vulva epidermoid carcinoma
(A431, ATCC, CRL-1555) and murine myeloma
P3-X63Ag8.653 (X63, ATCC, CRL-1580) were cultured in
DMEM: F12 (Life Technologies Inc., Grand Islan, NY)
supplemented with 10 % fetal bovine serum (FBS).
Lung metastasis murine models
Mice female of 6–8 week old female, were purchased from
the Center for Laboratory Animal Production
(CENPALAB, Havana, Cuba). Animal’s procedures were
performed in accordance with the guidelines stipulated by
Animal Subject Committee Review Board of the CIM and
CIM’’s Institutional Animal Care and Use Committees.
3LL-D122-metastasis model: C57BL/6 mice were injected
into lateral tail veins (i.v.) with 2.5 9 105 of tumor cells.
4T1-metastasis model: BALB/c mice were transplanted
subcutaneously (s.c). into the mammary gland with
1 9 104 of tumor cells. Primary tumor diameters were
measured every 2–3 days with a caliper and tumor volume
(mm3) was determined to t (...truncated)