Fig. 9

CCL3/VIRMA/SIRT1 pathway accelerates the ICC malignant process. a qRT-PCR analysis (up) and western blot analysis (down) of SIRT1 expression in rescue experiment. b Transwell migration assay and matrigel invasion assay of RBE cells and HuCCT1 cells in rescue experiment. Scale bars, 100 μm. c–f Intratumoral injections of CCL3 effectively promoted ICC subcutaneous tumor growth in nude mice, whereas the knockdown of VIRMA inhibited the progression. c Gross images of ICC subcutaneous tumor. d In vivo fluorescence imaging of nude mice bearing ICC tumors. e The tumor volume was monitored on daily basis, and tumor growth curves were generated. f The tumors were extracted and weighed after 4 weeks. g–j Stimulation of CCL3 effectively promoted ICC tumor liver metastasis in the orthotopic xenograft nude mice model, whereas the knockdown of VIRMA inhibited this progression. g Representative images of metastasis in the livers. h In vivo fluorescence imaging of orthotopic xenograft nude mouse model bearing ICC tumors. i The tumors were extracted and weighed after 8 weeks. j Quantification of the liver metastasis. k Sections of ICC subcutaneous tumor and l orthotopic xenograft tumor were stained with antibodies through IHC staining. The tumor tissues were stained with an antibody specific for VIRMA, SIRT1, or Ki67. Scale bars, 200 μm. *P < 0.05 **P < 0.01, ***P < 0.001