In vivo role of LINC00240 in gastric cancer

To examine the in vivo role of LINC00240, we inoculated subcutaneously a total of 6 × 10⁶ stably LINC00240-OE, LINC00240-KD or control HGC-27 cells into fossa axillaries of five-week-old female nude BALB/c mice (Vital River Laboratory, Beijing, China). Tumor growth was measured every two days as previously described [15 (link), 23 (link)]. To assess functions of LINC00240 during gastric cancer metastases in vivo, we injected a total of 2 × 10⁶ HGC-27 cells with stable LINC00240-KD (shNC, sh240-1, or sh240-2) or LINC00240-OE (NC or LINC00240) into tail vein of 5-week-old female nude BALB/c mice (Vital River Laboratory, Beijing, China) (n = 3 per group). Mouse lungs with metastatic tumors were formalin fixed, paraffin-embedded and stained with hematoxylin and eosin (HE). Immunohistochemistry (IHC) staining was performed in mouse livers with an antibody specific for vimentin as previously reported [24 (link), 25 (link)]. All procedures involving mice were approved by the Animal Care Committee of Shandong Cancer Hospital and Institute. All analyses were performed in a blinded fashion with individuals unaware of xenograft types.