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Outline
To investigate the effects of combined exposure of benzo[a]pyrene (B[a]P) and 2,2',4,4'-tetrabromobiphenyl ether (BDE-47), which widely coexist in the environment as endocrine-disrupting chemicals, on the proliferation, migration and invasion of mammary epithelial cells and their possible tumorigenic mechanisms.
Human mammary epithelial MCF-10A cells were treated with 1 μM B[a]P and gradient concentrations of BDE-47, alone or in combination. Cell cycle and apoptosis were detected by flow cytometry. Cell proliferation, migration and invasion were detected by CCK-8 method, scratch test and Transwell chamber. Protein expression levels of epithelial-to-mesenchymal transition(EMT) related markers and β-catenin were detected by Western blot.
Compared with control group, flow cytometry results showed that exposure to B[a]P and BDE-47 alone and in combination significantly decreased the proportion of G0/G1 cells and significantly increased the proportion of S and G2/M cells(P<0.05), but did not induce apoptosis in MCF-10A cells(P>0.05). The expression of cyclin-dependent kinase 4(CDK4) and CyclinD1 increased significantly(P<0.05), and CDK6 expression level had no significant difference (P>0.05). CCK-8 results showed that B[a]P and BDE-47, both alone and in combination, significantly enhanced cell viability, with the combined exposure producing a stronger effect(P<0.05). Scratch wound and Transwell assays demonstrated that both single and combined exposures enhanced cell migration and invasion(P<0.05), with the combined exposure group showing the most pronounced effects. Western blot analysis showed that EMT-related epithelial-associated proteins E-cadherin and ZO-1 were down-regulated (P<0.05), while mesenchymal associated proteins N-Cadherin, Vimentin and Snail were up-regulated (P<0.05), and the protein level of β-catenin was also increased(P<0.05), suggesting activation of the Wnt/β-catenin signaling pathway.
Combined exposure of B[a]P and BDE-47 promoted proliferation, migration and invasion of mammary epithelial cells, and the combined effect was stronger, which may induce EMT process of mammary epithelial cells through Wnt/β-catenin signaling pathway to play a tumorigenic role.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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