Event: 854

Key Event Title


Alterations, Cellular proliferation / hyperplasia

Short name


Alterations, Cellular proliferation / hyperplasia

Key Event Component


Process Object Action
cell proliferation Liver cell abnormal
hyperplasia hepatic oval stem cell increased
hyperplasia intrahepatic bile duct epithelial cell increased

Key Event Overview

AOPs Including This Key Event


AOP Name Role of event in AOP
Sustained AhR Activation leading to Rodent Liver Tumours KeyEvent



Level of Biological Organization


Biological Organization

Organ term


Organ term

Taxonomic Applicability


Term Scientific Term Evidence Link
rodentia rodentia NCBI

Life Stages


Sex Applicability


How This Key Event Works


Over the time period of high levels of sustained AHR activation, DLCs produce a complex pattern of cell proliferative responses. Teeguarden et al. (1999) observed that rats initiated with diethylnitrosamine (DEN) and then dosed with either 0.1 or 1 ng/ kg-d TCDD for one month exhibited a reduced labeling index relative to controls, and reduced BrdU-labeling was also observed following the 0.1 ng/kg-d TCDD dose after three months. Maronpot et al. (1993) observed a reduction in BrdU labeling index in hepatocytes at a low dose of 3.5 ng/kg-d TCDD in DEN-initiated rats after 30 weeks of TCDD administration but, at a dose of 125 ng/kg/d the labeling index was increased.

Both parenchymal calls and liver stem cells are likely involved in the organ-level response to sustained AHR activation. Early acti- vation of the AHR in zone 3 of the liver acinus causes decreased hepatocyte replication and may act as an indirect proliferative stimulus for stem cells and hepatoblasts (Andersen et al., 1997; Conolly and Andersen, 1997; Tritscher et al., 1992). Oval cells in the periportal region likely function as a source of replacement of hepatocytes after inhibition of normal hepatocyte replication- replacement (Paku et al., 2001; Sahin et al., 2008; Tanaka et al., 2011; Wang et al., 2003). While hepatocyte replication is considered the normal means for replacement of liver parenchyma, inhibition of hepatocyte replication in centrilobular regions induced by TCDD may induce normally quiescent liver stem cells to proliferate.

Following longer period of sustained AHR activation, organ- level increases in cell proliferation ensue, demonstrated by an in- crease in BrdU labeling and likely reflecting the regenerative response to organ-wide toxicity (Hailey et al., 2005).

Non-parenchymal cells, including stem cells, hepatoblasts, biliary cells, stellate cells, endothelial cells, and Kuppfer cells, play a role in this AOP. In rodents, TCDD elicits a fibrogenic and bile duct proliferative response that requires pathological alteration of stellate cell function and increased differentiation and growth of hepatoblasts and bile duct cells before 33 weeks of exposure. Retinoid depletion induces stellate cell proliferation, production of extracellular matrix components, and the transition to fibroblast; stellate cells maintain vitamin A homeostasis and respond to liver injury with formation of proliferative cytokines such as TGF-a and EGF (Friedman, 2008; Pintilie et al., 2010; Senoo et al., 2010). TCDD induces loss of retinoid content (presumably from stellate cells) and may disrupt the extensive communication between various liver cell types (Fletcher et al., 2001; Hoegberg et al., 2005; Pierre et al., 2014; Schmidt et al., 2003). Thus, TCDD-induced retinol loss from hepatic stellate cells may contribute to cell proliferation, biliary fibrosis, and cholangiolarcarcinoma (Fattore et al., 2000; Friedman, 2008; Hakansson and Hanberg, 1989; Schmidt et al., 2003).

AHR activation also induces changes in stem/oval cells. All of the rats receiving 100 ng/kg/day TCDD, the highest dose group animals in the 2-year cancer bioassay, developed oval cell hyperplasia with clear statistical increases in this endpoint at 22 ng/kg/day or greater (Hailey et al., 2005). Evidence points to the involvement of TNF-alpha regulation in the proliferative response of hepatic stem cells; this is likely mediated through modulation of the levels of TNF-alpha, altered beta-catenin signaling, and inhibition of cell-to-cell contact (Knight et al., 2000; Umannova et al., 2007; Vondracek et al., 2009; Dietrich et al., 2002; Prochazkova et al.,2011; Weiss et al., 2008). TNF-alpha is an inflammatory cytokine with an important role in liver tumor promotion. More research on how sustained AHR activation dysregulates normal TNF-alpha activity could be very impactful on evolving the AOP.

How It Is Measured or Detected


Bile duct hyperplasia and oval cell hyperplasia are measured histopathological observations using frequency of occurrence and a severity index.

Evidence Supporting Taxonomic Applicability


The proliferative response of the liver appears to occur in rodents but not humans.



Andersen, M.E., Birnbaum, L.S., Barton, H.A., Eklund, C.R., 1997. Regional hepatic CYP1A1 and CYP1A2 induction with 2,3,7,8-tetrachlorodibenzo-p-dioxin eval- uated with a multicompartment geometric model of hepatic zonation. Toxicol. Appl. Pharmacol. 144, 145-155.

Conolly, R.B., Andersen, M.E., 1997. Hepatic foci in rats after diethylnitrosamine initiation and 2,3,7,8-tetrachlorodibenzo-p-dioxin promotion: evaluation of a quantitative two-cell model and of CYP 1A1/1A2 as a dosimeter. Toxicol. Appl. Pharmacol. 146, 281-293.

Dietrich, C., Faust, D., Budt, S., Moskwa, M., Kunz, A., Bock, K.W., Oesch, F., 2002. 2,3,7,8-tetrachlorodibenzo-p-dioxin-dependent release from contact inhibition in WB-F344 cells: involvement of cyclin A. Toxicol. Appl. Pharmacol. 183, 117-126.

Fletcher, N., Hanberg, A., Håkansson, H., 2001. Hepatic vitamin a depletion is a sensitive marker of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure in four rodent species. Toxicol. Sci. 62, 166-175.

Hailey, J.R., Walker, N.J., Sells, D.M., Brix, A.E., Jokinen, M.P., Nyska, A., 2005. Clas- sification of proliferative hepatocellular lesions in harlan sprague-dawley rats chronically exposed to dioxin-like compounds. Toxicol. Pathol. 33, 165-174.

Hoegberg, P., Schmidt, C.K., Fletcher, N., Nilsson, C.B., Trossvik, C., Gerlienke Schuur, A., Brouwer, A., Nau, H., Ghyselinck, N.B., Chambon, P., Håkansson, H., 2005. Retinoid status and responsiveness to 2,3,7,8-tetrachlorodibenzo-p- dioxin (TCDD) in mice lacking retinoid binding protein or retinoid receptor forms. Chem. Biol. Interact. 156, 25-39.

Knight, B., Yeoh, G.C., Husk, K.L., Ly, T., Abraham, L.J., Yu, C., Rhim, J.A., Fausto, N., 2000. Impaired preneoplastic changes and liver tumor formation in tumor necrosis factor receptor type 1 knockout mice. J. Exp. Med. 192, 1809-1818.

Maronpot, R.R., Foley, J.F., Takahashi, K., Goldsworthy, T., Clark, G., Tritscher, A.,Portier, C., Lucier, G., 1993. Dose response for TCDD promotion of hep- atocarcinogenesis in rats initiated with DEN: histologic, biochemical, and cell proliferation endpoints 8. Environ. Heal. Perspect. 101, 634-642.

Paku, S., Schnur, J., Nagy, P., Thorgeirsson, S.S., 2001. Origin and structural evolution of the early proliferating oval cells in rat liver. Am. J. Pathol. 158, 1313-1323.

Pierre, S., Chevallier, A., Teixeira-Clerc, F., Ambolet-Camoit, A., Bui, L.-C., Bats, A.-S., Fournet, J.-C., Fernandez-Salguero, P., Aggerbeck, M., Lotersztajn, S., Barouki, R., Coumoul, X., 2014. Aryl hydrocarbon receptor-dependent induction of liver fibrosis by dioxin. Toxicol. Sci. 137, 114-124.

Pintilie, D.G., Shupe, T.D., Oh, S.-H., Salganik, S.V., Darwiche, H., Petersen, B.E., 2010. Hepatic stellate cells' involvement in progenitor-mediated liver regeneration. Lab. Invest. 90, 1199-1208.

Prochazkova, J., Kabatkova, M., Bryja, V., Umannova, L., Bernatík, O., Kozubík, A., Machala, M., Vondracek, J., 2011. The interplay of the aryl hydrocarbon receptor and b-catenin alters both AhR-dependent transcription and Wnt/b-catenin signaling in liver progenitors. Toxicol. Sci. 122, 349-360.

Sahin, M.B., Schwartz, R.E., Buckley, S.M., Heremans, Y., Chase, L., Hu, W.-S., Verfaillie, C.M., 2008. Isolation and characterization of a novel population of progenitor cells from unmanipulated rat liver. Liver Transpl. 14, 333-345.

Schmidt, C.K., Hoegberg, P., Fletcher, N., Nilsson, C.B., Trossvik, C., Hakansson, H., Nau, H., 2003. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) alters the endoge- nous metabolism of all-trans-retinoic acid in the rat. Arch. Toxicol. 77, 371-383.

Senoo, H., Yoshikawa, K., Morii, M., Miura, M., Imai, K., Mezaki, Y., 2010. Hepatic stellate cell (vitamin A-storing cell) and its relativeepast, present and future. Cell. Biol. Int. 34, 1247-1272.

Tanaka, M., Itoh, T., Tanimizu, N., Miyajima, A., 2011. Liver stem/progenitor cells: their characteristics and regulatory mechanisms. J. Biochem. 149, 231-239.

Teeguarden, J.G., Dragan, Y.P., Singh, J., Vaughan, J., Xu, Y.H., Goldsworthy, T., Pitot, H.C., 1999. Quantitative analysis of dose- and time-dependent promotion of four phenotypes of altered hepatic foci by 2,3,7,8-tetrachlorodibenzo-p- dioxin in female Sprague-Dawley rats. Toxicol. Sci. 51, 211-223.

Tritscher, A.M., Goldstein, J.A., Portier, C.J., McCoy, Z., Clark, G.C., Lucier, G.W., 1992. Dose-response relationships for chronic exposure to 2,3,7,8- tetrachlorodibenzo-p-dioxin in a rat tumor promotion model: quantification and immunolocalization of CYP1A1 and CYP1A2 in the liver. Cancer. Res. 52, 3436-3442.

Umannova, L., Zatloukalova, J., Machala, M., Krcmar, P., Majkova, Z., Hennig, B., Kozubík, A., Vondracek, J., 2007. Tumor necrosis factor-alpha modulates effects of aryl hydrocarbon receptor ligands on cell proliferation and expression of cytochrome P450 enzymes in rat liver “stem-like” cells. Toxicol. Sci. 99, 79-89.

Vondracek, J., Krcmar, P., Prochazkova, J., Trilecova, L., Gavelova, M., Skalova, L., Szotakova, B., Buncek, M., Radilova, H., Kozubik, A., Machala, M., 2009. The role of aryl hydrocarbon receptor in regulation of enzymes involved in metabolic activation of polycyclic aromatic hydrocarbons in a model of rat liver progenitor cells. Chem. Biol. Interact. 180, 226-237.

Wang, X., Foster, M., Al-Dhalimy, M., Lagasse, E., Finegold, M., Grompe, M., 2003. The origin and liver repopulating capacity of murine oval cells. Proc. Natl. Acad. Sci. U. S. A. 100 (Suppl. 1), 11881-11888.

Weiss, C., Faust, D., Schreck, I., Ruff, A., Farwerck, T., Melenberg, A., Schneider, S., Oesch-Bartlomowicz, B., Zatloukalova, J., Vondracek, J., Oesch, F., Dietrich, C., 2008. TCDD deregulates contact inhibition in rat liver oval cells via Ah receptor, JunD and cyclin A. Oncogene 27, 2198-2207.