Prediction of Prognosis in Breast Cancer by Using CD34 Immunostain and Quantitative Image Analysis
Ugur Ozerdem, Eva M Wojcik, Guliz A Barkan, Xiuzhen Duan, Cagatay Ersahin. Loyola University Medical Center, Chicago, IL
Background: Breast cancer is the most common cancer diagnosed in women. It is therefore imperative to establish new prognostic markers that can be easily utilized in breast pathology practice to stratify the patients. CD34 is expressed by both blood vessel endothelial cells and nascent (activated) lymphatic endothelial cells in tumors. CD34 immunostain is therefore in a unique position to identify angiogenesis and lymphangiogenesis simultaneously in a given tumor tissue. This investigation aims at establishing a quantitative analysis of CD34 expression in different stages of breast cancer tissues as a tangible prognostic tool in breast pathology.
Design: We used a tissue microarray, which contained 40 cores with stage IIA, IIB, IIIA, IIIB, and IIIC breast cancer, and 10 non-cancer breast cores. Tissue microarray slides were immunostained with CD34 for identification of blood vessel and nascent lymphatic endothelial cells. Immunostained slides were imaged with a high resolution digital camera. Digital images were analyzed using NIH ImageJ1.44 image analysis software. A CD34 score was quantified for each patient as a percentage of the CD34-positive microvessel area to the area of the microarray spot (CD34 area/area of entire tissue core). Statistical analyses were performed using Graphpad Prism Software. The scores in each position of tissue microarray were compared across each prognostic group. Bonferroni's multiple comparison test was used to compare prognostic groups in pairs.
Results: The mean CD34 score was 0.24%, 0.40%, 1.30%, 2.33%, 2.64%, and 3.44% for normal patients, patients with stage IIA, IIB, IIIA, IIIB, and IIIC breast cancer groups, respectively (p<0.0001). The mean CD34 score was 0.70% and 2.21% for lymph node-negative and lymph node–positive breast cancer groups, respectively (p<0.0001).
Conclusions: CD34 score of breast cancer, quantified by using CD34 immunostain and NIH ImageJ1.44 image analysis software, can predict prognosis. This method can be utilized as a practical and cost-effective alternative method to stratify breast cancer patients. Potentially, CD34 score can also be used to identify the patients who would benefit from targeted anti-blood vessel endothelial cell and anti-lymphatic endothelial cell therapies. Further studies are warranted.
Supported by a research grant from Loyola University Medical Center.
Wednesday, March 21, 2012 9:30 AM
Poster Session V # 29, Wednesday Morning