Effect of thyroid hormone responsive protein (THRP) expression on PC12 cell survival

The thyroid hormone responsive protein (THRP) is a novel gene product that remains responsive to thyroid hormone in the cerebral cortex of adult rats. The biological effects of THRP are currently unknown. Since thyroid hormones (TH) are known to cause cell death in primary neuronal cultures, the effect of exogenous THRP expression on PC12 cell viability was investigated. Co-transfection of the THRP expression plasmid with the selectable marker pSV2neo resulted in a lower number of surviving PC12 cells compared to transfection with pSV2neo and the empty vector, pSVL. Similar results were observed when PC12 cells were transfected with the plasmid pCMV. SPORT beta-gal with and without pSVL-THRP. However, expression of exogenous THRP in the colonic epithelial cell line Caco-2 and the glial cell line U251 had no effect on cell viability. Coexpression of THRP with either the wild-type (WT)-c-Abl or a kinase-defective mutant c-Abl (K290R) did not alter the cell viability changes induced by THRP alone. Under these experimental conditions the predominant form of cell death was necrosis as evidenced by in situ analyses, such as terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) and staining with membrane permeating and non-permeating nuclear dyes, Hoechst 33342 and propidium iodide respectively. In addition cell cycle arrest induced by THRP was demonstrated by reduced (3)H-thymidine incorporation into cellular DNA. The number of PC12 cells treated with 10(-7) M of l-3, 5, 3'-triiodothyronine (T(3)) was significantly reduced after the fourth day of culture. Treatment of the cells with T(3 )resulted in a dose dependent induction of THRP mRNA. It is concluded that: (1). THRP expression induces PC12 cell death; (2). under these experimental conditions the form of cell death is predominantly necrosis although cell cycle arrest may also occur; (3). the effect of THRP on cell viability is not modulated by c-Abl tyrosine kinase; and (4). the effect of T(3 )treatment on PC12 cell survival may be mediated by THRP.