Overexpression of clusterin in human breast carcinoma

Abstract

Clusterin has been implicated in numerous processes including active cell death, immune regulation, cell adhesion and morphological transformation. The purpose of this study was to examine clusterin expression in a large series of breast carcinomas by immunohistochemistry and in situ hybridization. The study included 40 samples of non-neoplastic glandular epithelia, 42 benign lesions, 15 atypical intraductal hyperplasias, 35 carcinomas in situ, 114 invasive carcinomas, and lymph node metastases from 40 patients. Epithelial normal cells were always negative for clusterin expression and only 19% of the benign lesions presented positive staining. In contrast to the benign lesions, however, the frequency of clusterin positive samples increased in atypical hyperplasias (47%, P = 0.08), intraductal carcinomas (49%, P = 0.01) and invasive carcinomas (53%, P < 0.001). Positive staining presented a cytoplasmic pattern, except in 3 cases of invasive carcinomas which had nuclear staining. Clusterin mRNA by in situ hybridization confirmed the specific cellular pattern of clusterin expression by immunohistochemistry. Clusterin expression was associated with large tumor size (P = 0.04), estrogen and progesterone receptor negative status (P = 0.02 and P = 0.001, respectively) and with the progression from primary carcinoma to metastatic carcinoma in lymph nodes (80% metastatic nodes had positive expression) (P = 0.004). Ten of 15 (67%) primary carcinomas without clusterin expression became positive in lymph node metastases, while most (22 of 25, 88%) of the clusterin-positive primary carcinomas were also immunoreactive in metastases. In survival analysis, clusterin expression did not represent a prognostic indicator by uni- or multivariate analysis. The increased clusterin expression in breast carcinomas tended to correlate inversely with the apoptotic index (P = 0.09) which indicates that clusterin gene expression is not a prerequisite to cellular death by apoptosis. From these results, we suggest that clusterin may have a role in tumorigenesis and progression of human breast carcinomas.

Figure 1.

Immunohistochemical analysis of clusterin expression in proliferative lesions and DCIS. A: Clusterin negative ductal hyperplasia without atypias. B: Atypical ductal hyperplasia showing positive staining. C and D: Strong cytoplasmic clusterin staining is seen in a low-grade DCIS with intermediate nuclear grade and absence of necrosis (C) and in a high-grade DCIS (D). All fields are magnified ×400. All sections were counterstained with hematoxylin.

Figure 2.

Positive cytoplasmic staining of clusterin in invasive breast carcinomas by immunohistochemistry and in situ hybridization. A and B: Immunoreactivity of clusterin in a lobular carcinoma (original magnifications: ×50 and ×400). C and D: In situ hybridization analysis of clusterin mRNA in poorly (C) and moderately (D) differentiated ductal carcinomas (both ×400). E and F: Lymph node metastasis of an infiltrating ductal carcinoma (×100 and ×400). All sections were counterstained with hematoxylin.