Purpose: Microarray studies have linked Annexin A8 RNA expression to a “basal cell–like” subset of breast cancers, including BRCA1-related cancers, that are characterized by
cytokeratin 5 ( CK5 ) and ...CK17 expression and show poor prognosis. We assessed Annexin A8's contribution to the overall prognosis and its expression in
normal, benign, and cancerous tissue and addressed Annexin A8's physiologic role in the mammary gland.
Experimental Design: Using microarrays and reverse transcription-PCR, the Annexin A8 expression was studied during mouse mammary gland development and in isolated mammary structures. Reverse transcription-PCR
on cultured human luminal and basal cells, along with immunocytochemistry on normal and benign breast tissues, was used for
cellular localization. Annexin A8's prognostic relevance and its coexpression with CK5 were assessed on tissue arrays of 1,631
cases of invasive breast cancer. Coexpression was further evaluated on a small cohort of 14 BRCA1-related breast cancers.
Results: Annexin A8 was up-regulated during mouse mammary gland involution and in pubertal ductal epithelium. Annexin A8 showed preferred expression in cultured basal cells but predominant luminal expression in normal human breast tissue in vivo . Hyperplasias and in situ carcinomas showed a strong staining of basal cells. Annexin A8 expression was significantly associated with grade ( P < 0.0001), CK5 ( P < 0.0001), and estrogen receptor status ( P < 0.0001); 85.7% BRCA1-related breast tumors coexpressed Annexin A8 and CK5.
Conclusion: Annexin A8 is involved in mouse mammary gland involution. In humans, it is a luminally expressed protein with basal expression
in cell culture and in hyperplasia/ductal carcinoma in situ . Expression in invasive breast carcinomas has a significant effect on survival ( P = 0.03) but is not independent of grade or CK5.
Analysis of cells is a cornerstone in biomedical research. Traditional methods for cell culture and tissue analysis can be replaced by various microchips as discussed in this chapter. A tissue array ...is an example of microchip that provides higher throughput of tissue analysis. Other microchips provide completely new functionalities for the researcher. One such example that will be discussed is cell culture chips. The cell culture vessel will be much more competent for studies of cell and will enable real-time monitoring of cell behavior and gene expression at a single cell level, have possibilities for studying chemotaxis and shear stress phenomena in a controlled manner, provide least tissue-like culture conditions by providing micro 3D structures, and provide arrays of integrated detectors. Tissue arrays and the simplest forms of cell culture chips are commercialized today, and we may expect a large activity in this field in the future.
Purpose
To investigate the possible clinical value of the expression of MMPs and their tissue inhibitors (TIMPs) by the different cellular types of the tumor scenario to predict biochemical ...recurrence in patients undergoing radical prostatectomy due clinically localized prostate cancer.
Methods
An immunohistochemical study was performed using tissue arrays and specific antibodies against MMPs-1, 2, 7, 9, 11, 13 and 14 and TIMPs-1, 2 and 3 on cancer specimens from 133 patients with clinical localized prostate carcinoma.
Results
Immunostaining for all the proteins studied was localized predominantly in tumor cells, but also in stromal cells in a significant percentage of prostate carcinomas, ranged from 20 to 50% for several proteins in fibroblast-like cells and in mononuclear inflammatory cells. Multivariate analysis according to a Cox model demonstrated that tumor stage (
P
< 0.0001) and Gleason grading (grades 7–10: 2.08 (1.1–3.9);
P
< 0.05) were significantly and independently associated with biochemical recurrence. Additionally, the expression of MMP-9 by fibroblast-like cells (
P
< 0.01) and MMP-13 by tumor cells (
P
< 0.05) were also variables significantly and independently associated with biochemical recurrence.
Conclusions
MMP-13 expression by tumor cells and MMP-9 by stromal fibroblast-like cells were independent factors of biochemical recurrence in prostate cancer.
Tissue arrays can evaluate molecular targets in high numbers of samples in parallel. Array construction presents technical difficulties and tissue arrayers are expensive, particularly for small and ...medium sized laboratories. This report describes a method for the construction of 36 sample arrays using widely available materials. A blunted 16 gauge needle for bone marrow aspiration was used to extract paraffin wax cylinders and manually define a 6 × 6 matrix on a blank paraffin wax block. Tissue cores from 36 paraffin wax embedded premalignant lesions and invasive cervical carcinomas were injected into the matrix using a 14 gauge needle. This tissue array was sectioned using a standard microtome and used for the immunodetection of CD44 variant 9 and interleukin 18 with satisfactory results. This method can be applied in any laboratory, without the need of specialised equipment, offering a good alternative for the wider application of tissue arrays.
High-throughput microarrays are being used in expression profiling analyses with the objectives of gene and pathway discovery, functional characterization of genes, and tumor subclassification. This ...review summarizes bladder cancer studies dealing with both in vitro models and clinical specimens, using distinct microarray platforms for target discovery.
Aims To investigate the expression of matrix metalloproteases (MMPs) and their inhibitors (TIMPs) in patients who develop local recurrence (LR) after mastectomy. Methods We analyzed the expressions ...of MMP-1, -2, -7, -9, -11, -13, -14, TIMP-1, -2, and -3, using immunohistochemical techniques, in primary tumors from patients without tumoral recurrence (n = 50), patients who developed distant metastasis (n = 50), and from patients who develop LRs (n = 25). LRs of the latter group were also analyzed for MMPs expression. All the patients underwent mastectomy. Results Score values for all MMPs and TIMPs were significantly higher in primary tumors of patients with distant metastasis. Primary tumors from patients with LR have lower expressions of MMPs and TIMPs compared with those from patients who developed distant metastasis, and with patients without recurrence for some MMPs. Remarkably, however, primary tumors from patients with LR showed significantly higher percentage of TIMP-1 and 2 expression in stromal cells compared to primary tumors from patients with distant metastasis or primary tumors from patients without tumoral progression. Furthermore, LRs had significantly higher MMP-9 expression than their corresponding primary tumors. Conclusions Our data indicate differences in MMPs/TIMPs expression between primary tumors of patients with LRs and of those with distant metastasis, both after mastectomy for breast cancer.
High avidity, pentameric, single-domain antibodies, oligomerized through the B subunit of verotoxin, are excellent immunohistochemical reagents. The resulting molecules are termed pentabodies. Here, ...we describe the immunostaining of tissue sections with ES1, a pentabody recognizing CEACAM6 which is overexpressed in several cancers. The advantages of pentabodies, compared to conventional antibody reagents, in immunohistochemical studies are highlighted.
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