This proposal is in response to RFA 94-001. According to the directives stated in the RFA, we will replicate the experiments reported by Goodman and Henderson that detected EMF induced general increases in gene expression. They will determine the ability of 60 HZ EMF to alter expression or activity of a panel of factors chosen for their association with cell proliferation and malignant transformation. Specificity of any changes will be assessed by comparison to EMF effects on housekeeping genes. In many early studies, the determination of biological activity was limited by the choice of malignantly transformed cells as targets for exposure to EMF. In order to assess EMF effects on a broad range of biological properties that relate to human health risks, we will replicate the original experimental conditions with a human endometrial cell based transformation assay system. The results obtained with these cells should directly relate to women's health issues. The human cell based assay system that we have established has the capability to detect alterations that influence cellular lifespan, that promote transformation of immortal cells of both stromal and epithelial lineage, and to detect alterations in stromal epithelial cell interactions. They propose to determine whether EMF changes the frequency at which cells escape from the limitations of finite life span to become immortal. To assess another potential biological effect of EMF, we will determine whether exposure to EMF causes transformation of SV40 immortalized endometrial stromal and epithelial cells. Concurrently, we will assess alterations in expression of cellular oncogenes, transcription factors, and enzymes that relate to malignant transformation.EMF induced changes will be related to cell lineage and stage of transformation. To improve the relevance of their results to women's health, we will assess EMF induced alterations of cellular factors that govern the cell's response to steroid hormones. If we identify EMF induced effects that relate to specific cytokines and transcription factors that are integral to the cell's response to hormones, we will attempt to prove the involvement of these factors in EMF effects by altering cells with specific genetic inhibitors. Because of the importance of stromal epithelial cell interactions, we will determine if exposure to EMF alters cellular interactions in ways that further neoplastic transformation. In order to improve their assessment of the human health risk associated with exposure to EMF, we will determine whether chronic exposure to EMF increases the incidence or degree of alterations in specific factors associated with transformation and determine the biological significance of these alterations by assessing concomitant changes to cellular behavior.
