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Usefulness of Immunohistochemistry for Microsatellite Instability Screening in Gastric Cancer.

Bae YS, Kim H, Noh SH, Kim H - Gut Liver (2015)

Bottom Line: Kaplan-Meier curves and a Cox proportional hazard regression model were used to conduct survival analyses.The MSI-H AGCs were correlated with older age (p<0.001), female gender (p=0.018), distal location (p<0.001), larger size (p=0.016), and intestinal type (p<0.001).Multivariate analysis revealed that the MSI-H phenotype was an independent favorable factor that was related to overall survival in patients with AGC (p<0.001).

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT

Background/aims: The usefulness of immunohistochemistry to screen for the microsatellite instability (MSI) phenotype in gastric cancer remains unclear. Moreover, the prognostic value of MSI phenotypes in gastric cancer has been debated.

Methods: The clinicopathologic parameters and survival outcomes of 203 MSI-high (MSI-H) and 261 microsatellite-stable (MSS) advanced gastric cancers (AGCs) were compared. Next, we compared the immunohistochemistry results for hMLH1 and hMSH2 with those of a polymerase chain reaction (PCR)-based method. Kaplan-Meier curves and a Cox proportional hazard regression model were used to conduct survival analyses.

Results: The MSI-H AGCs were correlated with older age (p<0.001), female gender (p=0.018), distal location (p<0.001), larger size (p=0.016), and intestinal type (p<0.001). Multivariate analysis revealed that the MSI-H phenotype was an independent favorable factor that was related to overall survival in patients with AGC (p<0.001). Compared with the PCR-based analysis, immunohistochemistry exhibited high sensitivity (91.1%) and specificity (98.5%) in the detection of MSI phenotypes.

Conclusions: MSI-H gastric cancers have distinct clinicopathologic features and better prognoses, which suggests the necessity of MSI analysis in gastric cancer. Immunohistochemistry can be a useful and reliable screening method in the assessment of MSI status in gastric cancer.

No MeSH data available.


Related in: MedlinePlus

Representative histology and immunohistochemistry results. A microsatellite instability-high (MSI-H) case (A) exhibiting the loss of the expression of hMLH1 (B) and the intact expression of hMSH2 (C). Stromal lymphocytes were used as an internal positive control. The intact expression of hMLH1 (E) and hMSH2 (F) was evident in a microsatellite stable case (D).
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f1-gnl-09-629: Representative histology and immunohistochemistry results. A microsatellite instability-high (MSI-H) case (A) exhibiting the loss of the expression of hMLH1 (B) and the intact expression of hMSH2 (C). Stromal lymphocytes were used as an internal positive control. The intact expression of hMLH1 (E) and hMSH2 (F) was evident in a microsatellite stable case (D).

Mentions: Paraffin-embedded tissue blocks were cut into 4 μm sections. IHC was performed using a Ventana XT automated stainer (Ventana Corp., Tucson, AZ, USA) with antibody to hMLH1 (ready-to-use, clone M1; Roche, Indianapolis, IN, USA) and hMSH2 (1:300, clone G219-1129; BD Pharmingen, San Jose, CA, USA). Sections were deparaffinized using EZ Prep solution (Ventana Corp.). CC1 standard (pH 8.4 buffer containing Tris/borate/EDTA) was used for antigen retrieval and blocked with inhibitor D (3% H2O2) for 4 minutes at 37°C. Slides were incubated with primary antibody for 40 minutes at 37°C followed by a universal secondary antibody for 20 minutes at 37°C. Slides were incubated in streptavidin-horseradish peroxidase (SA-HRP) D for 16 minutes at 37°C and then the substrate, 3,3′-diaminobenzidine tetrahydrochloride (DAB) H2O2, was added for 8 minutes followed by hematoxylin and bluing reagent counterstaining at 37°C. A loss of MMR protein expression was designated when none of the neoplastic epithelial cells showed nuclear staining, while normal expression was defined as the presence of nuclear staining of tumor cells, irrespective of the proportion or intensity (Fig. 1). Infiltrating lymphocytes, stromal cells and adjacent nonneoplastic epithelium served as internal positive controls. Two pathologists (Yoon Sung Bae and Hyunki Kim) assessed the IHC results without awareness of the MSI status of each case.


Usefulness of Immunohistochemistry for Microsatellite Instability Screening in Gastric Cancer.

Bae YS, Kim H, Noh SH, Kim H - Gut Liver (2015)

Representative histology and immunohistochemistry results. A microsatellite instability-high (MSI-H) case (A) exhibiting the loss of the expression of hMLH1 (B) and the intact expression of hMSH2 (C). Stromal lymphocytes were used as an internal positive control. The intact expression of hMLH1 (E) and hMSH2 (F) was evident in a microsatellite stable case (D).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4562780&req=5

f1-gnl-09-629: Representative histology and immunohistochemistry results. A microsatellite instability-high (MSI-H) case (A) exhibiting the loss of the expression of hMLH1 (B) and the intact expression of hMSH2 (C). Stromal lymphocytes were used as an internal positive control. The intact expression of hMLH1 (E) and hMSH2 (F) was evident in a microsatellite stable case (D).
Mentions: Paraffin-embedded tissue blocks were cut into 4 μm sections. IHC was performed using a Ventana XT automated stainer (Ventana Corp., Tucson, AZ, USA) with antibody to hMLH1 (ready-to-use, clone M1; Roche, Indianapolis, IN, USA) and hMSH2 (1:300, clone G219-1129; BD Pharmingen, San Jose, CA, USA). Sections were deparaffinized using EZ Prep solution (Ventana Corp.). CC1 standard (pH 8.4 buffer containing Tris/borate/EDTA) was used for antigen retrieval and blocked with inhibitor D (3% H2O2) for 4 minutes at 37°C. Slides were incubated with primary antibody for 40 minutes at 37°C followed by a universal secondary antibody for 20 minutes at 37°C. Slides were incubated in streptavidin-horseradish peroxidase (SA-HRP) D for 16 minutes at 37°C and then the substrate, 3,3′-diaminobenzidine tetrahydrochloride (DAB) H2O2, was added for 8 minutes followed by hematoxylin and bluing reagent counterstaining at 37°C. A loss of MMR protein expression was designated when none of the neoplastic epithelial cells showed nuclear staining, while normal expression was defined as the presence of nuclear staining of tumor cells, irrespective of the proportion or intensity (Fig. 1). Infiltrating lymphocytes, stromal cells and adjacent nonneoplastic epithelium served as internal positive controls. Two pathologists (Yoon Sung Bae and Hyunki Kim) assessed the IHC results without awareness of the MSI status of each case.

Bottom Line: Kaplan-Meier curves and a Cox proportional hazard regression model were used to conduct survival analyses.The MSI-H AGCs were correlated with older age (p<0.001), female gender (p=0.018), distal location (p<0.001), larger size (p=0.016), and intestinal type (p<0.001).Multivariate analysis revealed that the MSI-H phenotype was an independent favorable factor that was related to overall survival in patients with AGC (p<0.001).

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT

Background/aims: The usefulness of immunohistochemistry to screen for the microsatellite instability (MSI) phenotype in gastric cancer remains unclear. Moreover, the prognostic value of MSI phenotypes in gastric cancer has been debated.

Methods: The clinicopathologic parameters and survival outcomes of 203 MSI-high (MSI-H) and 261 microsatellite-stable (MSS) advanced gastric cancers (AGCs) were compared. Next, we compared the immunohistochemistry results for hMLH1 and hMSH2 with those of a polymerase chain reaction (PCR)-based method. Kaplan-Meier curves and a Cox proportional hazard regression model were used to conduct survival analyses.

Results: The MSI-H AGCs were correlated with older age (p<0.001), female gender (p=0.018), distal location (p<0.001), larger size (p=0.016), and intestinal type (p<0.001). Multivariate analysis revealed that the MSI-H phenotype was an independent favorable factor that was related to overall survival in patients with AGC (p<0.001). Compared with the PCR-based analysis, immunohistochemistry exhibited high sensitivity (91.1%) and specificity (98.5%) in the detection of MSI phenotypes.

Conclusions: MSI-H gastric cancers have distinct clinicopathologic features and better prognoses, which suggests the necessity of MSI analysis in gastric cancer. Immunohistochemistry can be a useful and reliable screening method in the assessment of MSI status in gastric cancer.

No MeSH data available.


Related in: MedlinePlus