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eBook details

• Title: Development of a Competitive Enzyme-Linked Immunosorbent Assay (ELISA) for Zeranol Detection and the Gene Regulation by Zeranol in Breast Cancer
• Author : Jieyu Liu
• Release Date : January 19, 2013
• Genre: Science & Nature,Books,Professional & Technical,Medical,
• Pages : * pages
• Size : 5748 KB

Description

Zeranol, an anabolic growth promoter, has been widely used in the livestock industry. Due to its potent estrogenic activity, Zeranol is recognized as a potential carcinogenic agent for breast cancer. To further study the association between breast cancer and Zeranol consumption, we attempted to develop a direct competitive enzyme-linked immunosorbent assay (ELISA) for the detection and quantitation of Zeranol residues in the clinical samples from normal humans and breast cancerpatients. Zearalanone, one of the major metabolites of Zeranol, was used as the hapten to produce the monoclonal antibody, anti-Zearalanone (anti-Z). The horseradish peroxidase-conjugated Zearalanone (HRP-Z) or HRP-conjugated Zeranol (HRP-z) was used to compete the limited binding sites of anti-Z with the Zeranol residues in samples. We have determined the optimal amount of anti-Z that can be placed on the plate, but pretty low signal was generated when HRP-Z was added to allow binding to anti-Z. The result suggests that anti-Z couldn’t recognize HRP-conjugated antigen for some reasons. In current study, the gene regulation of protein disulfide isomerase family A, member 3 (PDIA3) and estrogen receptor alpha (ESR1) by Zeranol has been investigated. Both in vitro and in vivo studies have shown that PDIA3 mRNA level was up-regulated but ESR1 was down-regulated in response to Zeranol. In the large-scale examination of human breast specimens, the same expression pattern was identified in primary cultured human breast cancer epithelial cells. ESR1 expression was negatively associated with the age; however, neither PDIA3 nor ESR1 was correlated with the pathological characteristics. The negative correlation between PDIA3 and ESR1 expression was found in cancerpatients and certain pathological characteristics including high graded stage, non triple-negativebreast cancer, peritumor lymphovascular invasion, estrogen receptor alpha positive, human epidermal growth factor receptor 2 negative, progesterone receptor positive, and T2 stage.

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