Utilization of Gene Profiling and Proteomics to Determine Mineral Pathogenicity in a Human Mesothelial Cell Line (LP9/TERT-1)

Abstract

Background-Identifying and understanding the early molecular events that underscore mineral pathogenicity using in vitro screening tests is imperative, especially given the large number of synthetic and natural fibers and particles being introduced into the environment. The purpose of the work described here was to examine the ability of gene profiling (Affymetrix microarrays) to predict the pathogenicity of various materials in a human mesothelial cell line (LP9/TERT-1) exposed to equal surface area concentrations (15×10 6 or 75×10 6 μm 2 /cm 2 ) of crocidolite asbestos, nonfibrous talc, fine titanium dioxide (TiO 2 ), or glass beads for 8 or 24 h. Since crocidolite asbestos caused the greatest number of alterations in gene expression, multiplex analysis (Bio-Plex) of proteins released from LP9/TERT-1 cells exposed to crocidolite asbestos was also assessed to reveal if this approach might also be explored in future assays comparing various mineral types. To verify that LP9/TERT-1 cells were more sensitive than other cell types to asbestos, human ovarian epithelial cells (IOSE) were also utilized in microarray studies. Upon assessing changes in gene expression via microarrays, principal component analysis (PCA) of these data was used to identify patterns of differential gene expression.

Induction of altered mRNA expression profiles caused by fibrous and granular dust

“…Our study and recent reports in the literature demonstrate that gene expression profiling in human lung epithelial cells can be an important tool for analyzing the pathogenicity of potentially harmful fibres and particles (42)(43)(44). Gene expression profiling, for example in response to asbestos, is valuable to define early molecular effects as demonstrated in diverse human cells, such as normal human bronchial epithelial cells (NHEC) (45), human lung adenocarcinoma cells (A549) (46,47), SV40-transformed human bronchial epithelial cells (BEAS-2B) and SV40-immortalized pleural mesothelial cells (MET5A) (47).…”

“…Gene expression profiling, for example in response to asbestos, is valuable to define early molecular effects as demonstrated in diverse human cells, such as normal human bronchial epithelial cells (NHEC) (45), human lung adenocarcinoma cells (A549) (46,47), SV40-transformed human bronchial epithelial cells (BEAS-2B) and SV40-immortalized pleural mesothelial cells (MET5A) (47). Changes in gene expression are also valuable to determine the pathogenicity pathway of asbestos fibres, as demonstrated in the human mesothelial (LP9/TERT-1) cell line (42).…”

The Role of Inflammation in Development and Therapy of Malignant Mesothelioma

“…We recently reported, using Affymetrix microarray and BioPlex analysis in primary and telomerase immortalized human mesothelial cells, that asbestos exposure causes increased inflammatory responses that include IL-1β, IL-13, basic Fibroblast Growth Factor (bFGF), VEGF and granulocyte colony stimulating factor (G-CSF) which may be responsible for mesotheliomagenic transformation of these cells (Shukla et al, 2009;Hillegass et al, 2010a). In addition, our in vivo work confirms that MM development in intraperitoneal mouse models is preceded by increased levels of many of these cytokines and growth factors (Hillegass et al, 2010b).…”

Secreted primary human malignant mesothelioma exosome signature reflects oncogenic cargo

“…Exposure of human MM cells to asbestos has been shown to facilitate autocrine production and transcriptional regulation of cytokines1920. Such findings support a malignant secretory network that can regulate the MM tumour microenvironment and fundamental to understanding the progression of various malignancies, including mesothelioma.…”