Fig. 5: Prospective study for proving that humanized molecular complex in pre-metastatic lungs trapped human cancer cells.

a Scheme of hSAAs-hCitFbg complex-induced metastasis. First, PBS, hFbg, or hCitFbg was injected into humanized hSAAs-Tg/mSAAs-KO mice. Then, labeled human cancer cells were injected. b Detection of SAAs proteins in lung tissues after injecting hCitFbg into hSAAs/mSAAs-/- mice by western blot analysis. The mSAAs-/-mice were used as a negative control. Two independent experiments. c Numbers of metastatic MCF7 (n = 4 hSAAs/mSAAs-/-; n = 3 mSAAs-/-) and MDAMB231 cells (n = 4 hSAAs/mSAAs-/--PBS; n = 7 hSAAs/mSAAs-/--hFbg and -hCitFbg; n = 7 SAAs-/-) in PBS-, hFbg-, and hCitFbg- treated-hSAAs/mSAAs-/- and mSAAs-/-mice. One-way ANOVA with Bonferroni correction. Data were mean ± SEM. d Representative IHC photos of metastatic MCF7 cells trapped by the hSAAs-hCitFbg complex. Scale bars, 20 μm. e Distances among hCitFbg, hSAA1/2, hSAA4, and MCF7 cells shown in (d) were analyzed (n = 4 hSAAs/mSAAs-/-; n = 3 mSAAs-/-) One-way ANOVA with Bonferroni correction. Data were mean ± SEM. f Summary of the distance among metastatic tumor cells, SAA3-hCitFbg, and hSAA1/2-hCitFbg. (n = 5 LLC-C57BL/6; n = 5 E0771-C57BL/6; n = 4 LLC-Scid; n = 3; MCF7- hSAAs/mSAAs-/-; n = 4 MDAMB231- hSAAs/mSAAs-/-) Data were mean ± SEM (Student’s two-sided t-test). g Similar strategy to Fig. 5a in hSAAs-Tg/mSAAs-KO in Rag1-/- background (upper left). Representative staining of Ki67 in lung 3 weeks after injection of tumor cells in hSAAs/mSAAs-/-/Rag1-/- and mSAAs-/-/Rag1-/- mice (upper right). Quantification of metastatic surface area in lungs 3 weeks after application of MCF7 (n = 5 mSAA-/-hSAAs-Tg/-Rag1-/-; n = 4 mSAA-/-hSAAs-/-Rag1-/-) and MDAMB231 (n = 4 mSAA-/-hSAAs-Tg/-Rag1-/-; n = 4 mSAA-/-hSAAs-/-Rag1-/-) cells (lower). Data were mean ± SEM (Student’s two-sided t-test).