Top

Published in:

01-10-2019 | Original Article

Immunohistochemical Assessment of the Expression of Biliary Transportation Proteins MRP2 and MRP3 in Hepatocellular Carcinoma and in Cholangiocarcinoma

Authors: Cinthya Santos Cirqueira, Aloisio Sousa Felipe-Silva, Alda Wakamatsu, Lidiane Vieira Marins, Eziel Cavalcanti Rocha, Evandro Sobroza de Mello, Venâncio Avancini Ferreira Alves

Published in: Pathology & Oncology Research | Issue 4/2019

Abstract

Multidrug resistance-associated protein 2 (MRP2) is a multi-specific organic anion transporter predominantly expressed in the canalicular membrane of hepatocytes, epithelial cells from gallbladder and apical membranes of proximal tubular kidney epithelium whereas multidrug resistance-associated protein 3 (MRP3) is present in the basolateral membrane of hepatocytes and cholangiocytes. This study aims to detect the expression of these transporters in hepatocellular carcinoma (HCC) and in cholangiocarcinoma (CC), searching for evidences for future studies on differential diagnosis and on clinical essays. The immunohistochemical reactivity (IHC) of these transporters was assessed in tissue microarrays of 80 HCC and 56 CC cases using monoclonal antibodies and compared with anatomopathological (AP) variables. The positivity of MRP2 was observed in 92.3% of HCC and in 96.3% of CC. The detection of high MRP2 expression in HCC was not significantly different (p > 0.05) according to the size, number of nodules architectural pattern and growth pattern of HCC and CC. Regarding histological grades, 22/22 well moderately differentiated HCC versus 50/56 poorly differentiated HCC were positive for MRP2. A trend for lower expression in poor differentiation HCC was found. And 50/50 well/moderately differentiated CC versus 2/4 poorly/undifferentiated CC were positive for MRP2. This result showed a reduced expression (p = 0,0004) in poorly differentiated CC. MRP3 positivity was observed in 18.8% of HCC and was not significantly different according to AP parameters. MRP3 was expressed in 44.5% CC, with a trend for lower expression in less differentiated CC and significantly lower rates in the ductular histological subtype (p = 0.023). The high expression of MRP2 in HCC and in CC is conserved regardless most of the anatomopathological parameters, except for a trend of lower expression in less differentiated HCC and CC. The observation of lower MRP3 expression in less differentiated CC and, especially, in the histological subtype with expression of hepatic progenitor cell phenotypes leads to future opportunities to evaluate the expression of this marker in cholangiocarcinomas.

Vasiliou V, Vasiliou K, Nebert DW (2009) Human ATP-binding cassette (ABC) transporter family. Hum Genomics 3:281–290. https://doi.org/10.1186/1479-7364-3-3-281 CrossRefPubMedPubMedCentral

Stefková J, Poledne R, Hubácek JA (2004) ATP-binding cassette (ABC) transporters in human metabolism and diseases. Physiol Res 53(3):235–243PubMed

Tarling EJ, de Aguiar Vallim TQ, Edwards PA (2013) Role of ABC transporters in lipid transport and human disease. Trends Endocrinol Metab 24(7):342–350. https://doi.org/10.1016/j.tem.2013.01.006 CrossRefPubMedPubMedCentral

Lagana SM, Salomao M, Remotti HE, Knisely AS, Moreira RK (2015) Bile salt export pump: a sensitive and specific immunohistochemical marker of hepatocellular carcinoma. Histopathology 66(4):598–602. https://doi.org/10.1111/his.12601 CrossRefPubMed

Nguyen T, Phillips D, Jain D, Torbenson M, Wu T-T, Yeh MM et al (2015) Comparison of 5 immunohistochemical markers of hepatocellular differentiation for the diagnosis of hepatocellular carcinoma. Arch Pathol Lab Med 139(8):1028–1034. https://doi.org/10.5858/arpa.2014-0479-OA CrossRefPubMed

Fujikura K, Yamasaki T, Otani K, Kanzawa M, Fukumoto T, Ku Y et al (2016) BSEP and MDR3 – useful immunohistochemical markers to discriminate hepatocellular carcinomas from intrahepatic cholangiocarcinomas and hepatoid carcinomas. Am J Surg Pathol 40(5):689–696. https://doi.org/10.1097/PAS.0000000000000585 CrossRefPubMed

Trauner M, Boyer JL (2003) Bile salt transporters: molecular characterization, function, and regulation. Physiol Rev 83(2):633–671. https://doi.org/10.1152/physrev.00027.2002 CrossRefPubMed

ABCC2: ATP-binding cassette, sub-family C (CFTR/MRP), member 2. [Internet]. The Human Protein Atlas. 2017. http://www.proteinatlas.org/ENSG00000023839-ABCC2/tissue. Accessed 31 Jan 2017

Taniguchi K, Wada M, Kohno K, Nakamura T, Kawabe T, Kawakami M et al (1996) A human canalicular multispecific organic anion transporter (cMOAT) gene is overexpressed in cisplatin-resistant human cancer cell lines with decreased drug accumulation. Cancer Res 56(18):4124–4129PubMed

10.

Chen Z, Shi T, Zhang L, Zhu P, Deng M, Huang C et al (2016) Mammalian drug efflux transporters of the ATP binding cassette (ABC) family in multidrug resistance: a review of the past decade. Cancer Lett 370(1):153–164. https://doi.org/10.1016/j.canlet.2015.10.010 CrossRefPubMed

11.

Nies AT, Konig J, Cui Y, Brom M, Spring H, Keppler D (2002) Structural requirements for the apical sorting of human multidrug resistance protein 2 (ABCC2). Eur J Biochem 269(7):1866–1876. https://doi.org/10.1007/s00424-006-0109-y CrossRefPubMed

12.

Jedlitschky G, Hoffmann U, Kroemer HK (2006) Structure and function of the MRP2 (ABCC2) protein and its role in drug disposition. Expert Opin Drug Metab Toxicol 2(3):351–366CrossRef

13.

Alrefai WA, Gill RK (2007) Bile acid transporters: structure, function, regulation and pathophysiological implications. Pharm Res 10:1803–1823. https://doi.org/10.1517/17425255.2.3.351 CrossRef

14.

Erlinger S, Arias IM, Dhumeaux D (2014) Inherited disorders of bilirubin transport and conjugation: new insights into molecular mechanisms and consequences. Gastroenterology 146(7):1625–1638. https://doi.org/10.1053/j.gastro.2014.03.047 CrossRefPubMed

15.

Keppler D (2014) The roles of MRP2, MRP3, OATP1B1, and OATP1B3 in conjugated hyperbilirubinemia. Drug Metab Dispos 42(4):561–565. https://doi.org/10.1124/dmd.113.055772 CrossRefPubMed

16.

Kool M, van der Linden M, de Haas M, Scheffer GL, de Vree JM, Smith AJ et al (1999) MRP3, an organic anion transporter able to transport anti-cancer drugs. Proc Natl Acad Sci U S A 96(12):6914–6919. https://doi.org/10.1073/pnas.96.12.6914 CrossRefPubMedPubMedCentral

17.

Scheffer GL, Kool M, Heijn M, de Haas M, Pijnenborg AC, Wijnholds J et al (2000) Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies. Cancer Res 60(18):5269–5277PubMed

18.

Klaassen CD, Aleksunes LM (2010) Xenobiotic, bile acid, and cholesterol transporters: function and regulation. Pharmacol Rev 62(1):1–96. https://doi.org/10.1124/pr.109.002014 CrossRefPubMedPubMedCentral

19.

ABCC3: ATP-binding cassette, sub-family C (CFTR/MRP), member 3 [Internet]. The Human Protein Atlas. 2017. http://www.proteinatlas.org/ENSG00000108846-ABCC3/tissue. Accessed 31 Jan 2017

20.

ABCC3: ATP-binding cassette, sub-family C (CFTR/MRP), member 3 [Internet]. GenAtlas - Universite Paris Descartes. 2017. http://genatlas.medecine.univ-paris5.fr/fiche.php?symbol=ABCC3. Accessed 31 Jan 2017

21.

König J, Rost D, Cui Y, Keppler D (1999) Characterization of the human multidrug resistance protein isoform MRP3 localized to the basolateral hepatocyte membrane. Hepatology 29(4):1156–1163. https://doi.org/10.1002/hep.510290404 CrossRefPubMed

22.

Ballatori N, Hammond CL, Cunningham JB, Krance SM, Marchan R (2005) Molecular mechanisms of reduced glutathione transport: role of the MRP/CFTR/ABCC and OATP/SLC21A families of membrane proteins. Toxicol Appl Pharmacol 204(3):238–255. https://doi.org/10.1016/j.taap.2004.09.008 CrossRefPubMed

23.

Belinsky MG, Dawson PA, Shchaveleva I, Bain LJ, Wang R, Ling V et al (2005) Analysis of the in vivo functions of Mrp3. Mol Pharmacol 68(1):160–168. https://doi.org/10.1124/mol.104.010587 CrossRefPubMed

24.

Halilbasic E, Claudel T, Trauner M (2013) Bile acid transporters and regulatory nuclear receptors in the liver and beyond. J Hepatol 58(1):155–168. https://doi.org/10.1016/j.jhep.2012.08.002 CrossRefPubMedPubMedCentral

25.

Sodani K, Patel A, Kathawala RJ, Chen Z-S (2012) Multidrug resistance associated proteins in multidrug resistance. Chin J Cancer 31(2):58–72. https://doi.org/10.5732/cjc.011.10329 CrossRefPubMedPubMedCentral

26.

Rau S, Autschbach F, Riedel HD, Konig J, Kulaksiz H, Stiehl A et al (2008) Expression of the multidrug resistance proteins MRP2 and MRP3 in human cholangiocellular carcinomas. Eur J Clin Investig 38(2):134–142. https://doi.org/10.1111/j.1365-2362.2007.01916.x CrossRef

27.

Tomonari T, Takeishi S, Taniguchi T, Tanaka T, Tanaka H, Fujimoto S et al (2016) MRP3 as a novel resistance factor for sorafenib in hepatocellular carcinoma. Oncotarget 7(6):7207–7215. https://doi.org/10.18632/oncotarget.6889 CrossRefPubMedPubMedCentral

28.

Zollner G, Wagner M, Fickert P, Silbert D, Fuchsbichler A, Zatloukal K et al (2005) Hepatobiliary transporter expression in human hepatocellular carcinoma. Liver Int (2):367–379. https://doi.org/10.1111/j.1478-3231.2005.01033.x CrossRef

29.

Nies AT, König J, Pfannschmidt M, Klar E, Hofmann WJ, Keppler D (2001) Expression of the multidrug resistance proteins MRP2 and MRP3 in human hepatocellular carcinoma. Int J Cancer 94(4):492–499. https://doi.org/10.1111/j.1478-3231.2005.01033.x CrossRefPubMed

30.

Borght VS, Libbrecht L, Blokzijl H, Faber NK, Moshage H, Aerts R et al (2005) Diagnostic and pathogenetic implications of the expression of hepatic transporters in focal lesions occurring in normal liver. J Pathol 207(4):471–482. https://doi.org/10.1002/path.1852 CrossRef

31.

Apte U, Krisnamurthy P (2010) Molecular pathology of liver diseases. In: Monga SPS (ed) Molecular pathology of liver diseases 1st edn. Springer, Boston, pp 147–164

32.

Korita PV, Wakai T, Shirai Y, Matsuda Y, Sakata J, Takamura M et al (2010) Multidrug resistance-associated protein 2 determines the efficacy of cisplatin in patients with hepatocellular carcinoma. Oncol Rep 4:965–972. https://doi.org/10.3892/ijmm.2012.1173 CrossRef

33.

Atilano-Roque A, Roda G, Fogueri U, Kiser JJ, Joy MS (2016) Effect of disease pathologies on transporter expression and function. J Clin Pharmacol 56(Suppl7):S205–S221. https://doi.org/10.1002/jcph.768 CrossRefPubMed

34.

Buettner S, van Vugt JLA, IJzermans J, Groot Koerkamp B (2017) Intrahepatic cholangiocarcinoma: current perspectives. Onco Targets Ther 10:1131–1142. https://doi.org/10.2147/OTT.S93629 CrossRefPubMedPubMedCentral

35.

Nakanuma Y, Kakuda Y (2015) Pathologic classification of cholangiocarcinoma: new concepts. Best Pract Res Clin Gastroenterol 29(2):277–293. https://doi.org/10.1016/j.bpg.2015.02.006 CrossRefPubMed

Title: Immunohistochemical Assessment of the Expression of Biliary Transportation Proteins MRP2 and MRP3 in Hepatocellular Carcinoma and in Cholangiocarcinoma
Authors: Cinthya Santos Cirqueira
Aloisio Sousa Felipe-Silva
Alda Wakamatsu
Lidiane Vieira Marins
Eziel Cavalcanti Rocha
Evandro Sobroza de Mello
Venâncio Avancini Ferreira Alves
Publication date: 01-10-2019
Publisher: Springer Netherlands
Published in: Pathology & Oncology Research / Issue 4/2019
Print ISSN: 1219-4956
Electronic ISSN: 1532-2807
DOI: https://doi.org/10.1007/s12253-018-0386-8

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 4/2019

Identification of Potential miRNAs Biomarkers for High-Grade Prostate Cancer by Integrated Bioinformatics Analysis

The Role of Testin in Human Cancers

Tumor Infiltrating Lymphocytes in Breast Cancer Patients with Progressive Disease during Neoadjuvant Chemotherapy

Association of Genetic Variants in NF-kB with Susceptibility to Breast Cancer: a Case Control Study

C-MYC, HIF-1α, ERG, TKT, and GSTP1: an Axis in Prostate Cancer?

Promoter DNA Methylation and mRNA Expression Level of p16 Gene in Oral Squamous Cell Carcinoma: Correlation with Clinicopathological Characteristics

Keynote webinar | Spotlight on antibody–drug conjugates in cancer