Laboratory of Molecular Toxicology, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509, USA.
Mol Carcinog. 2013 Jul;52(7):544-54. doi: 10.1002/mc.21889. Epub 2012 Mar 2.
The AhR was initially identified as a ligand-activated transcription factor mediating effects of chlorinated dioxins and polycyclic aromatic hydrocarbons on cytochrome P450 1 (CYP1) expression. Recently, evidence supporting involvement of the AhR in cell-cycle regulation and tumorigenesis has been presented. To further define the roles of the AhR in cancer, we investigated the effects of AhR expression on cell proliferation, migration, invasion, and tumorigenesis of MCF-7 human breast cancer cells. In these studies, the properties of MCF-7 cells were compared with those of two MCF-7-derived sublines: AH(R100) , which express minimal AhR, and AhR(exp) , which overexpress AhR. Quantitative PCR, Western immunoblots, 17β-estradiol (E2 ) metabolism assays, and ethoxyresorufin O-deethylase assays showed the lack of AhR expression and AhR-regulated CYP1 expression in AH(R100) cells, and enhanced AhR and CYP1 expression in AhR(exp) cells. In the presence of 1 nM E2 , rates of cell proliferation of the three cell lines showed an inverse correlation with the levels of AhR mRNA. In comparison with MCF-7 and AhR(exp) cells, AH(R100) cells produced more colonies in soft agar and showed enhanced migration and invasion in chamber assays with E2 as the chemoattractant. Despite the lack of significant AhR expression, AH(R100) cells retained the ability to form tumors in severe combined immunodeficient mice when supplemented with E2 , producing mean tumor volumes comparable to those observed with MCF-7 cells. These studies indicate that, while CYP1 expression and inducibility are highly dependent on AhR expression, the proliferation, invasion, migration, anchorage-independent growth, and estrogen-stimulated tumor formation of MCF-7 cells do not require the AhR.
AHR 最初被鉴定为一种配体激活的转录因子,介导氯化二恶英和多环芳烃对细胞色素 P4501(CYP1)表达的影响。最近,有证据表明 AHR 参与细胞周期调控和肿瘤发生。为了进一步确定 AHR 在癌症中的作用,我们研究了 AHR 表达对 MCF-7 人乳腺癌细胞增殖、迁移、侵袭和肿瘤形成的影响。在这些研究中,比较了 MCF-7 细胞与两种 MCF-7 衍生亚系的特性:表达最小 AHR 的 AH(R100)和过表达 AHR 的 AhR(exp)。定量 PCR、Western 免疫印迹、17β-雌二醇(E2)代谢测定和乙氧基 RESO 脱乙基酶测定显示,AH(R100)细胞缺乏 AHR 表达和 AHR 调节的 CYP1 表达,而 AhR(exp)细胞增强了 AHR 和 CYP1 表达。在 1nM E2 的存在下,三种细胞系的细胞增殖率与 AHR mRNA 水平呈反比。与 MCF-7 和 AhR(exp)细胞相比,AH(R100)细胞在软琼脂中产生更多的菌落,并在含有 E2 的腔室测定中显示出增强的迁移和侵袭能力,E2 作为趋化剂。尽管缺乏明显的 AHR 表达,但当补充 E2 时,AH(R100)细胞仍能在严重联合免疫缺陷小鼠中形成肿瘤,产生的平均肿瘤体积与 MCF-7 细胞观察到的体积相当。这些研究表明,虽然 CYP1 表达和诱导性高度依赖于 AHR 表达,但 MCF-7 细胞的增殖、侵袭、迁移、锚定非依赖性生长和雌激素刺激的肿瘤形成并不需要 AHR。