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. 2021 Mar 4;11(10):4770–4789. doi: 10.7150/thno.54235

Figure 7.

Figure 7

Inhibition of the CX3CL1/ICAM-1-mediated feedback cycle between circulating NSCLC cells and VBMECs by silencing CX3CR1 in A549 cells reduced NSCLC spinal metastasis and prolonged survival in mice. (A) Western blotting of CX3CR1 and LFA-1 protein levels in the indicated A549 cells. (B) Kaplan-Meier spinal metastasis-free curve of mice inoculated intracardially with control, CX3CR1-KD, CX3CR1-KD and Vector (CX3CR1-KD+Vector) or CX3CR1-KD and LFA-1-overexpression (CX3CR1-KD+LFA-1) A549 cells (n = 10). (C) Normalized BLI signals of spinal metastases. Data represent the mean ± standard deviation (SD) (n = 10). *P < 0.05 and NSP > 0.05. (D) Kaplan-Meier survival curve of mice. (E) Representative BLI, microCT, and histological (HE) images of bone lesions (B) invaded by tumors (T) in each group. Arrows indicate osteolytic bone lesions and dashed rectangles are used to mark the regions of interest for vertebral bone scan and analysis. (F) IHC images of p-PI3K, p-AKT, CX3CL1, and MMP-2 in tumor areas from serial sections of the indicated samples. (G, H) Quantification of osteolytic areas of spine and bone volume relative to total volume from microCT scans. Data represent the mean ± SEM (n = 5). **P < 0.01, ***P < 0.001, and NSP > 0.05. (I) mRNA levels of CX3CR1, LFA-1, Icam-1, and Gef-h1 in tumors of mice by RT-qPCR. Data represent the mean ± SEM (n = 10). *P < 0.05, **P < 0.01, ***P < 0.001, and NSP > 0.05.