Evidence suggests that estrogen affects the pulmonary response to carcinogenic pollutants, such as dioxins. In this study, we examined dioxin and estrogen signaling cross-talk in lung adenocarcinoma cell lines that were engineered to exhibit different aryl hydrocarbon receptor (AhR)/estrogen receptor (ER) α phenotypes. Data showed that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) weakly antagonized estrogen-activated ERα activity in cells expressing abundant ERα, but little AhR. Increase of AhR expression or presence of a dioxin-responsive element in proximity silenced the antiestrogenic effect of TCDD. AhR was bound to dioxin-responsive element and transcriptionally active in both TCDD-untreated and -treated lung adenocarcinoma cells. 17β-estradiol (E2) reduced basal and TCDD-induced AhR activity only in ERα-positive cells. AhR and ERα exhibited a protein-protein interaction in the presence of E2. Cotreatment with TCDD moderated this protein interaction. Colocalization of ERα and AhR at the estrogen-responsive site under E2 and TCDD/E2 treatments implied that E2 ∣ ERα might hijack AhR away from the dioxin-responsive site. Increasing the relative expression of AhR to ERα counteracted inhibition of AhR activity by E2 ∣ ERα. When AhR and ERα were both highly expressed, TCDD and E2 up-regulated expression of dual-responsive genes cytochrome P450 (CYP) 1A1 and CYP1B1 in a cumulative manner, increasing the danger of metabolic activation of carcinogens. Whereas TCDD ∣ AhR and E2 ∣ ERα appeared to regulate CYP1B1 separately through their binding sites, E2 ∣ ERα increased the TCDD responsiveness and mRNA expression of CYP1A1 in a noncanonical way. In conclusion, AhR/ERα expression pattern, estrogen level, and promoter context determine the genomic action of dioxin in lung adenocarcinoma cells.