Date of Award

2020

Document Type

Dissertation

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Pharmaceutical and Chemical Sciences

First Advisor

William K. Chan

First Committee Member

John C. Livesey

Second Committee Member

Miki S. Park

Third Committee Member

Wade A. Russu

Fourth Committee Member

Craig Vierra

Abstract

The aryl hydrocarbon receptor (AHR) is a ligand-activated signaling molecule which controls tumor growth and metastasis, T cell differentiation, and liver development. Expression levels of this receptor protein are sensitive to the cellular p23 protein levels in immortalized cancer cell lines. As little as 30% reduction of the p23 cellular content can suppress the AHR function. Here we reported that down-regulation of the p23 protein content in normal, untransformed human bronchial/tracheal epithelial cells to 48% of its content also suppresses the AHR protein levels to 54% of its content. This p23-mediated suppression of AHR is responsible for the repression of (1) the ligand-dependent induction of the cyp1a1 gene transcription; (2) the benzo[a]pyrene- or cigarette smoke condensate-induced CYP1A1 enzyme activity, and (3) the benzo[a]pyrene and cigarette smoke condensate-mediated production of reactive oxygen species. Reduction of the p23 content does not alter expression of oxidative stress genes or production of PGE2. Down-regulation of p23 suppresses the AHR protein levels in two other untransformed cell types, namely human breast MCF-10A and mouse immune regulatory Tr1 cells. Collectively, down-regulation of p23 suppresses the AHR protein levels in normal and untransformed cells and can in principle protect our lung epithelial cells from AHR-dependent oxidative damage caused by exposure to agents from environment and cigarette smoking.

The AHR is expressed in triple-negative and non-triple-negative breast cancer cells. It affects breast cancer growth and crosstalk with the estrogen receptor signaling. Normally the AHR is degraded shortly after ligand activation via the action of 26S proteasome. Here we report that the piperazinylpyrimidine compound Q18 triggers AHR protein degradation which is mediated through chaperone-mediated autophagy in triple-negative breast cancer cells (MDA-MB-468 and MDA-MB-231). This lysosomal degradation of AHR exhibits the following characteristics: (1) not observed in non-triple-negative breast cancer cells (MCF-7, T47D, and MDA-MB-361); (2) inhibited by progesterone receptor B but not estrogen receptor alpha; (3) reversed by chloroquine but not MG132; (4) required LAMP2A; (5) triggered by 6 amino-nicotinamide and starvation and (6) involved AHR-LAMP2A interaction mediated by 6 amino-nicotinamide and starvation. The NEKFF sequence localized at amino acid 558 of human AHR is a KFERQ-like motif of chaperone-mediated autophagy, essential for the LAMP2A-mediated AHR protein degradation.

Pages

123

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