Research Paper Volume 13, Issue 15 pp 19375—19396

THBS4/integrin α2 axis mediates BM-MSCs to promote angiogenesis in gastric cancer associated with chronic Helicobacter pylori infection

THBS4 mediates BM-MSC-induced promotion of GC angiogenesis in vivo. (A, B) Four-week-old nude male mice were injected with either SGC cells only or SGC and BM-MSCs transfected with recombinant lentiviral vectors. Three weeks following injection, the mice were euthanized, and the tumors were analyzed. n= 3 mice in control group and 5 mice per group in other groups. (C) Growth curve of tumors described in A. (D) Tumor weight of injected tumors at end-point. (E) H&E staining and Ki-67 and CD31 immunostaining of tumors described in A. (F, G) Quantification of staining presented in E performed with Image Plus. (H, I) Following AngioSense 750EX injection, tumor vascular permeability fluorescence imaging in vivo was performed to detect the neovascularization density of tumors in A. (J, K) Representative images and quantification of the CAM assay. n= 5 eggs in each group. (L, M) Western blot analysis of THBS4 in gastric tissues after NC BM-MSC or THBS4-knockdown BM-MSC transplantation. n= 3 mice in each group. (N) IF of THBS4 in gastric tissues after NC BM-MSC or THBS4-knockdown BM-MSC transplantation. (O) IF of CD31 and NG2 in gastric tissues in NC BM-MSC-transplanted mice and THBS4-knockdown BM-MSC-transplanted mice. n= 10 mice in each group.

Figure 6. THBS4 mediates BM-MSC-induced promotion of GC angiogenesis in vivo. (A, B) Four-week-old nude male mice were injected with either SGC cells only or SGC and BM-MSCs transfected with recombinant lentiviral vectors. Three weeks following injection, the mice were euthanized, and the tumors were analyzed. n= 3 mice in control group and 5 mice per group in other groups. (C) Growth curve of tumors described in A. (D) Tumor weight of injected tumors at end-point. (E) H&E staining and Ki-67 and CD31 immunostaining of tumors described in A. (F, G) Quantification of staining presented in E performed with Image Plus. (H, I) Following AngioSense 750EX injection, tumor vascular permeability fluorescence imaging in vivo was performed to detect the neovascularization density of tumors in A. (J, K) Representative images and quantification of the CAM assay. n= 5 eggs in each group. (L, M) Western blot analysis of THBS4 in gastric tissues after NC BM-MSC or THBS4-knockdown BM-MSC transplantation. n= 3 mice in each group. (N) IF of THBS4 in gastric tissues after NC BM-MSC or THBS4-knockdown BM-MSC transplantation. (O) IF of CD31 and NG2 in gastric tissues in NC BM-MSC-transplanted mice and THBS4-knockdown BM-MSC-transplanted mice. n= 10 mice in each group.