Zeranol Induces Cell Proliferation and Protein Disulfide Isomerase Expression in Mammary Gland of ACI Rat

Background: Zeranol is a non-steroidal anabolic growth promoter with potent estrogenic activity that is widely used as a growth promoter in the US beef industry. Consumption of beef derived from Zeranol-implanted cattle may be a risk factor for breast cancer. Protein disulfide isomerase (PDI) has been studied extensively as a key enzyme involving in the formation of the correct pattern of disulfide bonds in newly synthesized proteins. The relationship between PDI expression and cancer development has attracted interest of cancer researchers in recent years.

Materials And Methods: We implanted ACI rats with 12 mg Zeranol pellet and harvested the mammary tissues and tumor at day 110 after implantation and investigated the effect of Zeranol-implantation on cell proliferation by histological examination and proliferation in vitro. We also evaluated PDI mRNA expression in primary epithelial cells isolated from normal mammary glands and primary tumor cells from tumor specimens using real-time RT-PCR. To further confirm, we also evaluated the effect of Zeranol on PDI mRNA expression in primary epithelial cells isolated from normal mammary gland of ACI rats.

Results: We observed a palpable mammary tumor in one of three Zeranol-implanted ACI rats at day-110 post Zeranol-implantation. Zeranol-implantation significantly promoted the cell proliferation of primary mammary epithelial and stromal cells isolated from the mammary gland of normal ACI rats. PDI mRNA is over-expressed in primary tumor cells isolated from the tumor specimen and in Zeranol-treated primary cultured epithelial cells from the mammary gland of normal ACI rats.

Conclusion: These findings suggest that up-regulated expression of PDI may play a critical role in mammary tumorigenesis and cell proliferation in response to Zeranol. Our findings implicate PDI as a biomarker for mammary tumorigenesis.