Microsatellite Instability Occurs Rarely in Patients with Cholangiocarcinoma: A Retrospective Study from a German Tertiary Care Hospital - PubMed (original) (raw)

Microsatellite Instability Occurs Rarely in Patients with Cholangiocarcinoma: A Retrospective Study from a German Tertiary Care Hospital

Ria Winkelmann et al. Int J Mol Sci. 2018.

Abstract

Immune-modulating therapy is a promising therapy for patients with cholangiocarcinoma (CCA). Microsatellite instability (MSI) might be a favorable predictor for treatment response, but comprehensive data on the prevalence of MSI in CCA are missing. The aim of the current study was to determine the prevalence of MSI in a German tertiary care hospital. Formalin-fixed paraffin-embedded tissue samples, obtained in the study period from 2007 to 2015 from patients with CCA undergoing surgical resection with curative intention at Johann Wolfgang Goethe University hospital, were examined. All samples were investigated immunohistochemically for the presence of MSI (expression of MLH1, PMS2, MSH2, and MSH6) as well as by pentaplex polymerase chain reaction for five quasimonomorphic mononucleotide repeats (BAT-25, BAT-26, NR-21, NR-22, and NR-24). In total, 102 patients were included, presenting intrahepatic (n = 35, 34.3%), perihilar (n = 42, 41.2%), and distal CCA (n = 25, 24.5%). In the immunohistochemical analysis, no loss of expression of DNA repair enzymes was observed. In the PCR-based analysis, one out of 102 patients was found to be MSI-high and one out of 102 was found to be MSI-low. Thus, MSI seems to appear rarely in CCA in Germany. This should be considered when planning immune-modulating therapy trials for patients with CCA.

Keywords: PCR; cholangiocarcinoma; immunohistochemistry; microsatellite instability.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1

Figure 1

Kaplan–Meier graphs of the study population. (A) Survival data for moderate (group 1: UICC stadium I and II) and advanced (UICC stadium III and IV) cholangiocarcinoma (CCA) patients. (B) Survival data according to CCA location in the biliary tree (1: intrahepatic CCA, 2: perihilar CCA, 3: distal CCA).

Figure 2

Figure 2

Extrahepatic cholangiocarcinoma staining. (A): Hematoxylin and eosin (HE) staining. Samples immunohistochemically positive for (B): PMS2, (C): MLH1, (D): MSH6, (E): MSH2.

Figure 3

Figure 3

Typical allelic profiles of BAT-25, BAT-26, NR-21, NR-22, and NR-24. Only the range from 92 to 142 bp of the electropherograms comprising the region of interest is shown. The red line indicates the GeneScan 350 ROXTM dye Size Standard labeled with ROX displaying the 100 bp and 139 bp peak. The first column shows all traces, the second column shows the fluorescein (FAM)-labeled products (BAT-25, NR-21, and NR-22) depicted in blue. In the third column, the ATTO55O-labeled BAT-26 is separately displayed in black. HEX-labeled NR-24 is shown separately in green in column 4, and the fifth column shows the size standard in red. The expected sizes for the products NR-21, BAT-25, BAT-26, NR-24, and NR-22 were 99, 123, 124, 128, and 139 bp, respectively. The top panel shows the normal germline alleles with typical stutter bands. In the lower panels, the electropherograms of the only microsatellite instability (MSI)-high tumor sample (ID 105), the only MSI-low tumor sample (ID 121), and one MSI-stable tumor sample (ID 63) is shown. The arrows indicate the presence of shifted alleles. The asterisk depicts artifacts coming from bleed-trough from the FAM signal into the other signals, identifiable by peaks in the other colors occurring at the exact same size as the FAM peaks.

Figure 4

Figure 4

Representative stainings of the patients with MSI-low CCA (Pat ID 121, A–D) and MSI-high CCA (Pat ID 105, F–H). In both patients, no loss of expression was found for PMS2 (A,E), MLH2 (B,F), MLH1 (C,G), and MSH6 (D,H). Magnification: 40×, scale bar: 75 μm.

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