Activation, AhR leads to Increased, Invasion

Due to the extensive robust and concordant literature of the link between activation of the AhR-increased cell motility-increased invasion-breast cancer progression, the confidence in these key events was rated as high. However, due to the use of ligands to activate the AhR, it cannot be completely ruled out that alternative pathways (independent of the AhR) can also contribute to these features. For instance, 2 main pathways seem to explain this increase in migration and invasion: the c-Src/HER1/STAT5b, and ERK1/2 pathways. Yet, these pathways seem only to explain the relation between the AhR activation and cell migration / invasion, when the ligand used is hexachlorobenzene, an organochlorinated pesticide (Pontillo et al., 2011 Apr, Miret et al., 2016 Jul, Pontillo et al., 2013 May 1). Even though alternative mechanisms may present themselves, all studies blocked the AhR pathway and found a decrease in cell migration/invasion. The evidence for alternative mechanisms was therefore classified as “moderate” and the biological plausibility of KER was also classified as “moderate”.

The activation of the AhR through the use of different ligands (benzophenone, butyl benzyl phthalate, di-n-butyl phthalate, hexachlorobenzene, chlorpyrifos, TCDD) or the blockage of the AhR (silencing, KO or antagonism) increased or decreased cell invasion, respectively (Parks et al., 2014 Nov, Qin et al., 2011 Oct 20, Nguyen et al., 2016 Nov 15, Miret et al., 2016 Jul, Shan et al., 2020 Nov, Narasimhan et al., 2018 May 7, Hsieh et al., 2012 Feb, Pontillo et al., 2013 May 1, Miller et al., 2005, Belguise et al., 2007 Dec 15, Yamashita et al., 2018 May 1, Miret et al., 2020 May). The dose–response concordance for cell invasion was demonstrated using increasing doses of hexachlorobenzene, benzo[a]pyrene, chlorpyrifos and TCDD (Miret et al., 2016 Jul, Shan et al., 2020 Nov, Pontillo et al., 2013 May 1, Miller et al., 2005, Miret et al., 2020 May). To further explore cell invasion, Nguyen et al. created a model of a lymphatic barrier using a three-dimensional lymph endothelial cell as a monolayer co-cultured with spheroids of MDA-MB231 cells (Nguyen et al., 2016 Nov 15). They found that silencing or antagonizing the AhR (DIM) or activating the AhR (FICZ) respectively decreased or increased invasion of the lymphatic barrier.

On an organ level, in vivo, an increase in metastasis has been found in mice and zebrafish after the activation of the AhR with different ligands (butyl benzyl phthalate, di-n-butyl phthalate, hexachlorobenzene, TCDD) (Goode et al., 2014, Shan et al., 2020 Nov, Narasimhan et al., 2018 May 7, Hsieh et al., 2012 Feb, Pontillo et al., 2013 May 1). In the zebrafish model, Narasimham et al. treated the animals either with triple negative MDA-MB-231 cells only (untreated) or with MDA-MB-231 cells treated with an AhR inhibitor (CB7993113 or CH22319) (Narasimhan et al., 2018 May 7). Untreated fish had significantly more metastasis (OR = 9, IC95%=3–35). Similar results were found using mice models (Goode et al., 2014, Shan et al., 2020 Nov, Narasimhan et al., 2018 May 7, Hsieh et al., 2012 Feb, Pontillo et al., 2013 May 1).