To determine the role of ERβ in 25-HC-mediated LAC metastasis in vivo, we constructed LAC metastatic model by intravenously injecting with A549 cells or A549-depleting ERβ, followed by injecting with 0 or 0.085 μM 25-HC via tail vein every 2days.The results showed that 25-HC increased the number and size of tumor nodules, suggesting that 25-HC accelerated LAC metastasis, which was significantly reduced by ERβ knockdown (Fig. 7A and B).Consistently, HE staining also verified that 25-HCenhanced the number of tumor nodules, which was blocked by ERβ knockdown (Fig. 7C). Here, ESR2 is linked to neoplasm.