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Expression of Tight Junction Molecules in Breast Carcinomas Analysed by Array PCR and Immunohistochemistry

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Pathology & Oncology Research

Abstract

In the past few decades an enormous amount of data became known to clarify the molecular composition and architecture of tight junctions (TJs). Despite the efforts, the expression and function of several TJ genes and proteins in breast carcinoma are still not known and some of the data are contradictory. The expression of forty-four TJ associated genes was examined at mRNA level in eighteen invasive ductal breast carcinoma samples and corresponding normal breast tissues by using low density array PCR. Expressions of claudins (CLDNs) 5, 10, 16, 17, and 18, and ZO-1, ZO-2 were evaluated by immunohistochemistry as well. Using immunohistochemical phenotype as a surrogate for the genetic subtype, 11 luminal A, 3 luminal B, 3 triple negative and one HER2+ cases were included. Ten genes were significantly downregulated in tumors compared with normal breast tissues (CLDNs 5, 10, 16, 18, 19, CTNNAL1, JAM-B, ZO-1, ZO-2 and PARD3), whereas one gene (CLDN17) was significantly up-regulated in tumors when compared with normal breast. At protein level CLDNs 5, 10, 16, 18, ZO-1 and ZO-2 were downregulated in tumors as compared with normal breast tissue. CLDN17 showed variable expression in tumor tissues in comparison to normal breast. In the single HER2+ tumor when compared with the other subtypes CLDNs 5, 16, 17, 18, CTNNAL1, JAM-B, ZO-1, ZO-2 and PARD3 genes were found to be upregulated. We found altered TJ genes and proteins whose expression has not yet been associated with breast carcinoma. Our findings show a tendency of TJ genes and proteins to be downregulated in breast cancer. Further studies are necessary to examine whether the downregulation of the above mentioned TJ associated genes and proteins may contribute to the malignant progression of invasive ductal breast carcinomas.

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Abbreviations

AJ-:

Adherens junctions

BSA-:

Bovine serum albumine

CDC42-:

Cell division cycle 42 (GTP binding protein, 25 kDa)

CGN-:

Cingulin

CLDN-:

Claudin

CLDN1-:

Claudin 1

CLDN10-:

Claudin 10

CLDN11-:

Claudin 11 (oligodendrocyte transmembrane protein)

CLDN12-:

Claudin 12

CLDN14-:

Claudin 14

CLDN15-:

Claudin 15

CLDN16-:

Claudin 16

CLDN17-:

Claudin 17

CLDN18-:

Claudin 18

CLDN19-:

Claudin 19

CLDN2-:

Claudin 2

CLDN20-:

Claudin 20

CLDN3-:

Claudin 3

CLDN4-:

Claudin 4

CLDN5-:

Claudin 5

CLDN6-:

Claudin 6

CLDN7-:

Claudin 7

CLDN8-:

Claudin 8

CLDN9-:

Claudin 9

CLDND2-:

Claudin domain containing 2

CRB3-:

Crumbs homolog 3 (Drosophila)

CTNNAL1-:

Catenin (cadherin-associated protein), alpha-like 1

CTNNBIP1-:

Catenin, beta interacting protein 1

DAPI-:

4′-6-Diamidino-2-phenylindole’

F11R-:

F11 receptor

GAPDH-:

Glyceraldehyde-3-phosphate dehydrogenase

HPRT1-:

Hypoxanthine phosphoribosyltransferase 1

IDC-:

Invasive ductal carcinoma

JAM-B-:

Junctional adhesion molecule B

JAM-C-:

Junctional adhesion molecule C

MAGI1-:

Membrane associated guanylate kinase, WW and PDZ domain containing 1

MAGIX-:

MAGI family member, X-linked

MARK2-:

MAP/microtubule affinity-regulating kinase

MLLT4-:

Myeloid/lymphoid or mixed-lineage leukemia (trithorax homolog, Drosophila); translocated to, 4

MPDZ-:

Multiple PDZ domain protein

MPP5-:

Membrane protein, palmitoylated 5 (MAGUK p55 subfamily member 5)

OCLN-:

Occludin

PARD3-:

par-3 partitioning defective 3 homolog

PARD6A-:

par-6 partitioning defective 6 homolog alpha

PBS-:

Phosphate buffered saline

RHOA-:

Ras homolog gene family, member A

RPL13A-:

Ribosomal protein L13a

SDHA-:

Succinate dehydrogenase complex, subunit A, flavoprotein (Fp)

SYMPK-:

Symplekin

TGFB1-:

Transforming growth factor, beta 1

TJ-:

Tight junction

ZO-1-:

Tight junction protein 1 (zona occludens 1)

ZO-2-:

Tight junction protein 2 (zona occludens 2)

ZO-3-:

Tight junction protein 3 (zona occludens 3)

TLDA-:

TaqMan Low Density Array

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Acknowledgements

We thank Rigóné Káli Elvira for the careful reading of our manuscript and her valuable comments. This study was supported by grants: AVON-EAGC 2005, MKOT–GSK 2006, MKOT-GSK 2008, OTKA- 49559 (2005–2009), ETT-088-01/2009.

Competing interests

The author(s) declare that they have no financial or non-financial competing interests. They do not hold any patents relating to the content of this manuscript.

Authors’ contributions

A-MT and JK analysed the selected cases, evaluated the immunohistochemical results and wrote the article. AMSz helped in selecting the genes for TLDA and isolated RNA with A-MT. EJ carried out the immunohistochemical reactions. ZsS participated in the evaluation of the immunohistochemical reactions and in the writing of the manuscript in its final stage. LH, IAM, ZsB, IB and AZ performed surgery and provided samples. SF performed biopsy of specimens after surgery.

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Authors and Affiliations

  1. 2nd Department of Pathology, Semmelweis University, Ulloi ut 93, 1091, Budapest, Hungary

    Anna-Mária Tőkés, Attila Marcell Szász, Éva Juhász, Zsuzsa Schaff & Janina Kulka

  2. 1st Department of Surgery, Semmelweis University, Ulloi ut 78, 1091, Budapest, Hungary

    László Harsányi & István Arthur Molnár

  3. Department of Surgery and Vascular Surgery, Uzsoki Memorial Hospital, Uzsoki u. 29, 1145, Budapest, Hungary

    Zsolt Baranyai, István Besznyák Jr & Attila Zaránd

  4. Department of Pathology, Uzsoki Memorial Hospital, Uzsoki u. 29, 1145, Budapest, Hungary

    Ferenc Salamon

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  1. Anna-Mária Tőkés
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Correspondence to Anna-Mária Tőkés.

Electronic Supplementary Material

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ESM. 1

The relevant literature data on TJ gene and protein expression in breast carcinomas are presented in Supplementary Table 1. The primary and secondary antibodies used for TJ protein immunodetection are presented in Supplementary Table 2 (DOC 148 kb)

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Tőkés, AM., Szász, A.M., Juhász, É. et al. Expression of Tight Junction Molecules in Breast Carcinomas Analysed by Array PCR and Immunohistochemistry. Pathol. Oncol. Res. 18, 593–606 (2012). https://doi.org/10.1007/s12253-011-9481-9

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