Sulfatase 2 facilitates lymphangiogenesis in breast cancer by regulating VEGF-D

Abstract

In our previous studies, sulfatase 2 (Sulf2) was found to upregulate vascular endothelial growth factor-D (VEGF-D) expression in breast cancer. As VEGF-D plays an important role in lymphangiogenesis, we hypothesized that Sulf2 facilitates lymphangiogenesis in breast cancer by regulating VEGF-D. To evaluate the functions of Sulf2 on lymphangiogenesis in breast cancer, proliferation, apoptosis, cell cycle, cell mobility and tube-formation of lymphatic endothelial cells (LECs) were measured in vitro. Lymphangiogenesis in nude mouse ears and breast cancer xenografts were examined in vivo. Furthermore, the expression levels of related signaling pathway genes were screened and verified in LECs. We found that Sulf2 significantly increased the mobility and tube formation of the LECs, inhibited cisplatin-induced LEC apoptosis, but had no effect on cell proliferation and the cell cycle. Moreover, recombinant Sulf2 (rSulf2) combined with VEGF-D further promoted the proliferation, cell cycle, mobility and tube-like structure formation in the LECs, and at the same time inhibited cisplatin-induced apoptosis especially in the late stage. Sulf2 also significantly increased the density of lymphatic vessels in mouse ears and breast cancer xenografts in vivo. AKT1 was also shown to be upregulated and activated by Sulf2. Our results confirmed that Sulf2 facilitated lymphangiogenesis in breast cancer cells by regulating VEGF-D and that the AKT1-related signaling pathway was involved.

Isolation and characterisation of lymphatic endothelial cells from lung tissues affected by lymphangioleiomyomatosis

“…Though VEGF-D has been reported as a lymphangiogenic factor, we found that VEGF-D showed significant promotion only to migration, but had little effect on the proliferation of LECs. This is similar to the results in previous studies 26 , 27 . We think that VEGF-D appears to be more important in migration rather than proliferation of LECs.…”

Isolation and Characterisation of Lymphatic Endothelial Cells from Lung Tissues Affected by Lymphangioleiomyomatosis

“…Though VEGF-D has been reported as a lymphangiogenic factor, we found that VEGF-D showed signi cant promotion only to migration, but had little effect on the proliferation of LECs. This is similar to the results in previous studies 26,27 . We think that VEGF-D appears to be more important in migration rather than proliferation of LECs.…”

Release of HER2 repression of trefoil factor 3 (TFF3) expression mediates trastuzumab resistance in HER2+/ER+ mammary carcinoma

“…Western blot analysis was performed as previously described [ 29 ]. The primary antibodies used are as follows: phospho-AKT1 S473 (Abcam ab66138) [ 96 ], pan-AKT (Abcam ab8805) [ 97 ], β-Actin (Santa Cruz sc-47778), BCL-2 (Santa Cruz sc-509), phospho-cMET Y1234/1235 (Cell Signaling 3077S), cMET (Cell Signaling 8198S), phospho-cSRC Y416 (Santa Cruz sc-101802), cSRC (Santa Cruz sc-8056), phospho-EGFR Y1068 (Cell Signaling 3777S) [ 98 ], EGFR (Abcam ab2430) [ 99 ], phospho-p44/42 MAPK T202/Y204 (Cell Signaling 4370S) [ 100 ], p44/42 MAPK (Cell Signaling 4695S) [ 101 ], phospho-HER2/ neu Y1248 (Santa Cruz sc-12352-R) [ 60 ], HER2/ neu (Abcam ab2428) [ 102 ], phospho-HER3 Y1289 (Cell Signaling 4791S) [ 98 ], HER3 (Santa Cruz sc-415) [ 103 ], phospho-HER4 Y1284 (Cell Signaling 4757S) [ 104 ], HER4 (Santa Cruz sc-8050) [ 105 ], phospho-IGF1Rβ Y1135/1136 (Cell Signaling 3024S), IGF1Rβ (Cell Signaling 9750S), p27 kip1 (Cell Signaling 3686S), phospho-p65 S536 (Cell Signaling 3033S), p65 (Cell Signaling 4764S), phospho-STAT3 Y705 (Cell Signaling 9131S), STAT3 (Cell Signaling 9139S), and TFF3 (Abcam ab108599) [ 25 ]. The secondary anti-rabbit, anti-mouse and anti-goat horseradish peroxidase (HRP)-conjugated antibodies were obtained from Cell Signaling Technology.…”