Neuropilin-1 Interacts with Fibronectin-1 to Promote Epithelial–Mesenchymal Transition Progress in Gastric Cancer

Figures

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  Figure 1. NRP1 highly expressed in gastric cancer and predicts poor prognosis. (A–B) Survival curve of high NRP1 expression in patients with gastric cancer. (C) According to the TCGA database, high expression of NRP1 in gastric cancer tissues compared with normal tissues. (D) The high expression of NRP1 in metastatic gastric cancer. (E) Expression of NRP1 in gastric cancer patients with different pathological grade. (F) Expression of NRP1 in gastric cancer patients of different clinical stage. (G) Protein levels of NRP1 in ten pairs of fresh tumor tissues and adjacent normal tissues. Differences were considered significant at P < 0.05 and labeled with *.

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  Figure 2. (A) NRP1 enrichment gene and its correlated with EMT makers analysis. Go analysis enriched up-regulated gene function (B) Enrichment analysis of related pathway in NRP1 high expression samples in TCGA. (C–G) A Correlation analysis between NRP1 and EMT markers in gastric cancer.

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  Figure 3. Establishment of NRP1 overexpression and knockdown cell lines (A) Analysis of NRP1 expression in different cell lines using Western blot. (B) Detection of protein levels of NRP1 in gastric cancer cell lines after NRP1 overexpression of knockdown. (C) Wound healing was used to detect cell migration after ectopic alteration of NRP1 expression. (D) Transwell was performed to detect cell invasion after ectopic alteration of NRP1 expression. (E) Cell proliferation was analyzed by colony formation. (F) Protein levels of E-cadherin, N-cadherin and Vimentin were detected by Western blot. Differences were considered significant at P < 0.05 and labeled with *.

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  Figure 4. NRP1 promotes tumor growth in vivo. (A) Pictures of tumor-bearing animals after euthanasia. (B) Tumor volumes of each group. (C) Expression of NRP1 in solid tumors. (D–E) IHC was performed to detect the expression of E-cadherin and Vimentin. Differences were considered significant at P < 0.05 and labeled with *.

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  Figure 5. Ginsenoside Rg3 blocks the interaction of NRP1 and FN1. (A) Molecular docking and Co-IP conforming that Ginsenoside Rg3 blocks the interaction of NRP1 and FN1. (B) Transwell was performed to detect cell invasion after Ginsenoside Rg3 treatment. (C) Wound healing was used to detect cell migration after Ginsenoside Rg3 treatment. (D) Cell proliferation was analyzed by colony formation ability. Differences were considered significant at P < 0.05 and labeled with *.

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  Figure 6. Ginsenoside Rg3 inhibit gastric cancer cell progression in vivo. (A) Pictures of tumor-bearing animals and solid tumors after euthanasia in subcutaneous model. (B) Tumor volumes of each group. (C) Tumor metastasis after Ginsenoside Rg3 treatment. (D) Schematic diagram of this study. Differences were considered significant at P < 0.05 and labeled with *.