Perfluorooctanoic acid induces apoptosis through the p53-dependent mitochondrial pathway in human hepatic cells: a proteomic study.

Perfluorooctanoic acid (PFOA) is one of the most commonly used perfluorinated compounds, and exposure to it has been associated with a number of adverse health effects. However, the molecular mechanisms involved in PFOA toxicity are still not well characterized. In the present study, flow cytometry analysis revealed that PFOA induced oxidative stress, cell cycle arrest and apoptosis in human non-tumor hepatic cells (L-02). Furthermore, we investigated the alterations in protein profile within L-02 cells exposed to PFOA, aiming to explore the mechanisms underlying PFOA hepatotoxicity on the proteome level. Of the 28 proteins showing significant differential expression in response to PFOA, 24 were down-regulated and 4 were up-regulated. This proteomic study proposed that the inhibition of some proteins, including GRP78, HSP27, CTSD and hnRNPC may be involved in the activation of p53, which consequently triggered the apoptotic process in L-02 cells. Induction of apoptosis via the p53-dependent mitochondrial pathway is further suggested as one of the key toxicological events occurring in L-02 cells under PFOA stress. We hope these data will shed new light on the molecular mechanisms responsible for PFOA-mediated toxicity in human liver cells, and from such studies useful biomarkers indicative of PFOA exposure could be developed.

[1]  A. Tomida,et al.  Glucose‐regulated protein 78: A new partner of p53 in trophoblast , 2009, Proteomics.

[2]  N. Cherry,et al.  Perfluorinated acids and hypothyroxinemia in pregnant women. , 2011, Environmental research.

[3]  Lingling Sun,et al.  Glucose regulated protein 78 (GRP78) is overexpressed in colorectal carcinoma and regulates colorectal carcinoma cell growth and apoptosis. , 2011, Acta histochemica.

[4]  Heqing Shen,et al.  Perfluorooctanoic acid induces gene promoter hypermethylation of glutathione-S-transferase Pi in human liver L02 cells. , 2012, Toxicology.

[5]  A. Lampen,et al.  Perfluorooctanoic acid affects the activity of the hepatocyte nuclear factor 4 alpha (HNF4α). , 2012, Toxicology letters.

[6]  J. S. Ahn,et al.  Cathepsin D inhibits oxidative stress-induced cell death via activation of autophagy in cancer cells. , 2012, Cancer letters.

[7]  Kyle Christian,et al.  Interaction of Heterogeneous Nuclear Ribonucleoprotein C1/C2 with a Novel cis-Regulatory Element within p53 mRNA as a Response to Cytostatic Drug Treatment , 2008, Molecular Pharmacology.

[8]  C. Peters,et al.  Potential role for Cathepsin D in p53-dependent tumor suppression and chemosensitivity , 1998, Oncogene.

[9]  Z. Chang,et al.  Nuclear efflux of heterogeneous nuclear ribonucleoprotein C1/C2 in apoptotic cells: a novel nuclear export dependent on Rho-associated kinase activation , 2004, Journal of Cell Science.

[10]  M. Sousa,et al.  The protective role of yeast Cathepsin D in acetic acid‐induced apoptosis depends on ANT (Aac2p) but not on the voltage‐dependent channel (Por1p) , 2013, FEBS letters.

[11]  Nizar M. Mhaidat,et al.  Inhibition of GRP78 sensitizes colorectal cancer c ells to paclitaxelinduced apoptosis by activation of caspase4 , 2011 .

[12]  C. Epstein,et al.  The in vivo gene expression signature of oxidative stress. , 2008, Physiological genomics.

[13]  P. Lam,et al.  Differential expression of chicken hepatic genes responsive to PFOA and PFOS. , 2007, Toxicology.

[14]  Hwee Tong Tan,et al.  Membrane proteins and membrane proteomics , 2008, Proteomics.

[15]  J. Depierre,et al.  Sub-acute, moderate-dose, but not short-term, low-dose dietary pre-exposure of mice to perfluorooctanoate aggravates concanavalin A-induced hepatitis. , 2013, Toxicology letters.

[16]  Kathryn S Lilley,et al.  All about DIGE: quantification technology for differential-display 2D-gel proteomics , 2004, Expert review of proteomics.

[17]  K. Öllinger,et al.  Cathepsin D mediates cytochrome c release and caspase activation in human fibroblast apoptosis induced by staurosporine , 2003, Cell Death and Differentiation.

[18]  Marie Aune,et al.  Exposure of Perfluorinated Chemicals through Lactation: Levels of Matched Human Milk and Serum and a Temporal Trend, 1996–2004, in Sweden , 2006, Environmental health perspectives.

[19]  M E Hurtt,et al.  Mechanisms of extrahepatic tumor induction by peroxisome proliferators in male CD rats. , 2001, Toxicological sciences : an official journal of the Society of Toxicology.

[20]  M. Gorospe,et al.  Heterogeneous Nuclear Ribonucleoprotein C1/C2 Controls the Metastatic Potential of Glioblastoma by Regulating PDCD4 , 2012, Molecular and Cellular Biology.

[21]  Xiao-zhong Hu,et al.  Effects of perfluorooctanoate and perfluorooctane sulfonate exposure on hepatoma Hep G2 cells , 2009, Archives of Toxicology.

[22]  K. Roberg,et al.  Lysosomal release of cathepsin D precedes relocation of cytochrome c and loss of mitochondrial transmembrane potential during apoptosis induced by oxidative stress. , 1999, Free radical biology & medicine.

[23]  M. Murphy,et al.  Polymorphisms in the p53 pathway , 2006, Oncogene.

[24]  N. Mhaidat,et al.  Inhibition of GRP 78 sensitizes colorectal cancer cells to paclitaxel-induced apoptosis by activation of caspase-4 , 2011 .

[25]  L. Chan,et al.  Proteomic analysis of hepatic protein profiles in rare minnow (Gobiocypris rarus) exposed to perfluorooctanoic acid. , 2008, Journal of proteome research.

[26]  M. Rosen,et al.  Gene expression profiling in the lung and liver of PFOA-exposed mouse fetuses. , 2007, Toxicology.

[27]  Gloria B Post,et al.  Perfluorooctanoic acid (PFOA), an emerging drinking water contaminant: a critical review of recent literature. , 2012, Environmental research.

[28]  V. Laurent-Matha,et al.  Cathepsin D: newly discovered functions of a long-standing aspartic protease in cancer and apoptosis. , 2006, Cancer letters.

[29]  S. Lowe,et al.  Control of apoptosis by p53 , 2003, Oncogene.

[30]  G. Olsen,et al.  Perfluoroalkyl chemicals and human fetal development: an epidemiologic review with clinical and toxicological perspectives. , 2009, Reproductive toxicology.

[31]  D. Latchman,et al.  Do heat shock proteins have a role in breast cancer? , 1996, British Journal of Cancer.

[32]  D. Grandér,et al.  Reactive oxygen species and mitochondria mediate the induction of apoptosis in human hepatoma HepG2 cells by the rodent peroxisome proliferator and hepatocarcinogen, perfluorooctanoic acid. , 2001, Toxicology and applied pharmacology.

[33]  Zhuchu Chen,et al.  Identification of differential proteins in nasopharyngeal carcinoma cells with p53 silence by proteome analysis , 2007, FEBS letters.

[34]  Guido Kroemer,et al.  Mitochondrial control of cell death , 2000, Nature Medicine.

[35]  M. Cohen,et al.  Overexpression of GRP78 in complete hydatidiform moles. , 2012, Gynecologic oncology.

[36]  Ching-Hsein Chen,et al.  GRP78 Knockdown Enhances Apoptosis via the Down-Regulation of Oxidative Stress and Akt Pathway after Epirubicin Treatment in Colon Cancer DLD-1 Cells , 2012, PloS one.

[37]  Fan Wang,et al.  The hepatitis B virus X protein downregulates NF-κB signaling pathways through decreasing the Notch signaling pathway in HBx-transformed L02 cells. , 2013, International journal of oncology.

[38]  He-qing Huang,et al.  Alterations of protein profile in zebrafish liver cells exposed to methyl parathion: a membrane proteomics approach. , 2012, Chemosphere.

[39]  K. Yu,et al.  Induction of oxidative stress and apoptosis by PFOS and PFOA in primary cultured hepatocytes of freshwater tilapia (Oreochromis niloticus). , 2007, Aquatic toxicology.

[40]  Haiyan Zhou,et al.  The hepatitis B virus X protein downregulates NF-κ B signaling pathways through decreasing the Notch signaling pathway in HBx-transformed L 02 cells , 2013 .

[41]  Eugenia G. Giannopoulou,et al.  Pharmacoproteomic study of the natural product Ebenfuran III in DU-145 prostate cancer cells: the quantitative and temporal interrogation of chemically induced cell death at the protein level. , 2013, Journal of proteome research.

[42]  M. Candeias.  The can and can't dos of p53 RNA. , 2011, Biochimie.

[43]  M. Sherman,et al.  Hsp27 modulates p53 signaling and suppresses cellular senescence. , 2007, Cancer research.

[44]  Fatihah Suja,et al.  Contamination, bioaccumulation and toxic effects of perfluorinated chemicals (PFCs) in the water environment: a review paper. , 2009, Water science and technology : a journal of the International Association on Water Pollution Research.

[45]  L. Yin,et al.  Lysosomal and mitochondrial pathways in H2O2‐induced apoptosis of alveolar type II cells , 2005, Journal of cellular biochemistry.

[46]  J. Bertoglio,et al.  Lysosomal and mitochondrial pathways in miltefosine-induced apoptosis in U937 cells , 2007, Apoptosis.

[47]  A. Giwercman,et al.  Sperm DNA integrity in relation to exposure to environmental perfluoroalkyl substances - a study of spouses of pregnant women in three geographical regions. , 2012, Reproductive toxicology.

[48]  G. Kroemer,et al.  Heat shock proteins: endogenous modulators of apoptotic cell death. , 2001, Biochemical and biophysical research communications.

[49]  He-qing Huang,et al.  Differential expression profile of membrane proteins in zebrafish (Danio rerio) brain exposed to methyl parathion , 2011, Proteomics.

[50]  S H Kaufmann,et al.  Mammalian caspases: structure, activation, substrates, and functions during apoptosis. , 1999, Annual review of biochemistry.

[51]  Hongqin Zhuang,et al.  Down-regulation of HSP27 sensitizes TRAIL-resistant tumor cell to TRAIL-induced apoptosis. , 2010, Lung cancer.

[52]  S. Pizzo,et al.  GRP78: a chaperone with diverse roles beyond the endoplasmic reticulum. , 2008, Histology and histopathology.

[53]  Jiayin Dai,et al.  Proteomic analysis for testis of rats chronically exposed to perfluorododecanoic acid. , 2010, Toxicology letters.

[54]  Xiaofeng Yao,et al.  Genotoxic risk and oxidative DNA damage in HepG2 cells exposed to perfluorooctanoic acid. , 2005, Mutation research.