The Role of β-Arrestin1 in Esophageal Squamous Cell Carcinoma

Figures

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  Figure 1. β-arrestin1 is aberrantly upregulated in human ESCC tissues and cell lines and regulates the AKT/GSK3β/β-catenin signaling pathway. (A) Western Blot analysis of β-arrestin1 expression in tumor tissues and adjacent normal tissues in patients with ESCC. (B) Western Blot analysis of β-arrestin1 expression in the four ESCC cell lines TE-1, ECA-109, KYSE-410 and KYSE-520, and a normal esophageal epithelial cell line HEEC. (C) ECA-109 cells were transfected with sh-NC, sh-β-arrestin1-1 and sh-β-arrestin1-2, and the expression of β-arrestin1 was determined using Western blot. (D) ECA-109 cells were transfected with pcDNA-NC and pcDNA-β-arrestin1, and the expression of β-arrestin1 was determined using Western blot. (E) The protein expressions of p-AKT, AKT, p-GSK3β, GSK3β and β-catenin were measured by Western blotting assay. (F) Quantified evaluation of p-AKT, AKT, p-GSK3β, GSK3β and β-catenin. *P<0.05, **P<0.01, ***P<0.001 vs control group. #P<0.05, ###P<0.001 vs pcDNA-β-arrestin1 group.

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  Figure 2. β-arrestin1 regulates proliferation, migration, invasion and EMT of ESCC cells. (A) The cell proliferation ability was determined by Cell Counting kit-8 (CCK-8) assay. (B) The protein expressions of cyclinD1, CDK2 and p21 were measured by Western blotting assay. (C) Wound healing assays were performed to determine cell migration. (D) Quantitative results regarding the cell migration. (E) Trans-well assays were performed to determine cell invasion. (F) Quantitative results regarding the cell invasion. (G) The protein levels of MMP2 and MMP9 were measured by Western blotting assay. (H) The protein levels of E-cad, N-cad, vimentin and SANI1 were measured by Western blotting assay. *P<0.05, **P<0.01, ***P<0.001 vs control group. #P<0.05, ##P<0.01, ###P<0.001 vs pcDNA-β-arrestin1 group.

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  Figure 3. β-arrestin1 regulates tumor growth in vivo model of ESCC. (A) β-arrestin1 expression level in vivo model of ESCC after transfection with sh-NC, sh-β-arrestin1-1 and sh-β-arrestin1-2 was determined using Western blot. (B) β-arrestin1 expression level in vivo model of ESCC after transfection with pcDNA-NC and pcDNA-β-arrestin1 was determined using Western blot. (C) β-arrestin1 level was determined using immunohistochemistry assay. (D) Photographs of tumor tissues from different groups. (E) Growth curves of tumor volumes in xenografts of nude mice were determined based on tumor volume measured every 5 days. (F) Growth curves of tumor weights in xenografts of nude mice were determined based on tumor weight detected every 5 days. **P<0.01, ***P<0.001 vs control group. #P<0.05, ## P<0.01, ###P<0.001 vs pcDNA-β-arrestin1 group.

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  Figure 4. β-arrestin1 regulates tumor migration and invasion in vivo model of ESCC. (A) The protein levels of β-arrestin1, MMP2 and MMP9 were measured by Western blotting assay. (B) The protein expressions of E-cad, N-cad, vimentin and SANI1 were measured by Western blotting assay. *P<0.05, **P<0.01, ***P<0.001 vs control group. #P<0.05, ###P<0.001 vs pcDNA-β-arrestin1 group.