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. 2023 Apr 15;15(4):2656–2675.

Figure 4.

Figure 4

CD31 promotes DLBCL metastasis by activating AKT-OPN-EGFR pathway. A. We used immunohistochemical staining (IHC) to detect OPN expression in tissues and organs invaded by DLBCL cells (On the left). Con represent S4, S5 and S6 DLBCL patients’ samples. We also obtained samples from relapsed/refractory DLBCL patients by using needle aspiration and detected OPN by IHC (on the right). B. Western-blot was used to detect the effects of three groups (siRNA-CD31, GSK690693 and Odanacatib) on EXP-1 cells genes. DLBCL cells were treated with GSK690693 or Odanacatib (1 uM) for different hours. C. Three groups of EXP-1 cells were treated respectively with siRNA-CD31, GSK690693 or Odanacatib (1 uM) for 5 days. Then they were injected into three groups of C57BL/6Ly5.2 mice. Each group contains 10 mice. The number of invaded tissues were calculated in the 4th week. D. The survival curves of every group were analyzed. E. We detected the CD31 and OPN of 4 groups’ cells by using IF. Scale bar = 100 μm. The magnification is 200 times. F. 4 group of 3 × 107 lentiviral vectors induced CD31 overexpression DLBCL cells were injected into C57BL/6Ly5.2 mice through the tail vein. The Wright’ s staining was used to detected tissues infiltrated by CD31 DLBCL cells in the fourth week. Scale bar = 400 μm. The magnification is 50 times. Scale bar = 100 μm. The magnification is 200 times. G. HE staining was used to detect the structure of vessels in mice. The red circle shows DLBCL cells. Scale bar = 100 μm. The magnification is 400 times. H. TEM was used to observe the normal structure of endothelial cells. The red circles indicate the TJ that closed the spaces between endothelial cells. The magnification is shown in the figure. I. The abnormal TJ around the tumor tissue. As indicated by the arrows, the TJ went from being closed to being broken. J. DLBCL destroyed the structure of TJ. The arrow represents DLBCL cells moving to the opposite side. The red circles indicate complete TJ. K. The red circle above indicates split TJ. The red circles below indicate abnormal mitochondria. L. GSEA was used to detect EGFR-ZO-1/ZO-1 pathway. EGFR-IN-42 is an EGFR inhibitor. Real-time PCR and Western-blot were used to detect the expression ZO-1 and ZO-1. IHC: immunohistochemical staining; HE: hematoxylin-eosin staining; TEM: transmission electron microscope; TJ: tight junctions. All experiments were repeated at least in triplicate (*P < 0.05, **P < 0.01, ***P < 0.001).