Fig. 6

A After luciferase-labeled CAL-27 cells were constructed, the successful construction of the cell line was confirmed by luciferase fluorescence detection. B LCN2-overexpressing and LCN2-knockdown CAL-27-Luci cells were constructed by lentiviral transfection and confirmed by immunoblotting. C After a mouse tongue in situ tumor model was constructed, changes in mouse body weight were detected; the weight loss of the LCN2-OV group was greatest, and the weight loss of the shLCN2 group was lowest. D In situ tumors from the mouse tongue were dissected and weighed. The LCN2-OV group had the heaviest tumors, indicating that the tumors were the largest, while the shLCN2 group had the lightest tumors, indicating the smallest tumor volume. E After overexpression of LCN2, all mice with in situ tumors of the tongue developed lymph node metastasis (small dots in the submandibular region), while lymph node metastasis occurred in 1 mouse in the control group. (Scale bar: 1 cm). F After the expression of LCN2 was inhibited, all of the mice with in situ tumors of the tongue developed lymph node metastasis (small dots in the submandibular region), while lymph node metastasis occurred in 1 mouse in the control group. G Representative images of tongue tumors and cervical lymph nodes in the LCN2-ov group. HE staining showed tongue tumors and lymph node metastases in the LCN2-overexpressing group, and overexpression of LCN2 was confirmed by IHC staining. (Scale bar: 100 μm). H Representative images of tongue tumors and cervical lymph nodes in mice in the shLCN2 group. HE staining showed tongue tumors and lymph node metastases in the control group, but no metastases were found in the shLCN2 group, as detected by HE staining. The expression of LCN2 was clearly inhibited, as shown by IHC color development. (Scale bar: 100 μm)