Elevated FOXC2 Expression Promotes Invasion of HCC Cell Lines and is Associated with Poor Prognosis in Hepatocellular Carcinoma Cell Physiol Biochem 2017;44:99–109.

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Elevated FOXC2 Expression Promotes Invasion of HCC Cell Lines and is Associated with Poor Prognosis in Hepatocellular Carcinoma Cell Physiol Biochem 2017;44:99–109 - DOI:10.1159/000484586 Fig. 1. Evaluation of FOXC2 expression in primary human HCC tissues and HCC cell lines. (A) Analyses of FOXC2 mRNA expression in 84 pairs of HCC samples and adjacent normal liver tissue. (B) mRNA expression of FOXC2 in TNM I - II stage and TNM III stage patients by qRT-PCR. (C) Western blot analysis was performed to assess FOXC2 protein levels in HCC cell lines and LO2 cells. **P< 0.01. © 2017 The Author(s). Published by S. Karger AG, Basel - CC BY-NC-ND 4.0

Elevated FOXC2 Expression Promotes Invasion of HCC Cell Lines and is Associated with Poor Prognosis in Hepatocellular Carcinoma Cell Physiol Biochem 2017;44:99–109 - DOI:10.1159/000484586 Fig. 2. FOXC2 overexpression is associated with aggressive HCC. (A) Representative IHC staining of FOXC2 in adjacent normal tissues (left) and HCC tissues (right) (IHC ×200). (B) Kaplan-Meier curve indicating that patients with high expression of FOXC2 have a worse overall survival compared to patients with low expression of FOXC2 (P=0.0063). (C) Kaplan-Meier analyses indicating that patients in TNM III stage had a shorter overall survival (P=0.0048). © 2017 The Author(s). Published by S. Karger AG, Basel - CC BY-NC-ND 4.0

Elevated FOXC2 Expression Promotes Invasion of HCC Cell Lines and is Associated with Poor Prognosis in Hepatocellular Carcinoma Cell Physiol Biochem 2017;44:99–109 - DOI:10.1159/000484586 Fig. 3. Knockdown of FOXC2 expression inhibits HCC cell growth in vitro. (A) The protein level of FOXC2 was markedly reduced in MHCC-97H and Hep3B after silencing. (B) FOXC2 silencing significantly inhibits cell viability both in MHCC-97H and Hep3B cells. (C) Knockdown of FOXC2 induces cell apoptosis as measured by flow cytometry both in MHCC-97H and Hep3B cells. (D) Knockdown of FOXC2 inhibited colony formation as demonstrated by colony formation assay in MHCC-97H and Hep3B cells. **P < 0.01. © 2017 The Author(s). Published by S. Karger AG, Basel - CC BY-NC-ND 4.0

Elevated FOXC2 Expression Promotes Invasion of HCC Cell Lines and is Associated with Poor Prognosis in Hepatocellular Carcinoma Cell Physiol Biochem 2017;44:99–109 - DOI:10.1159/000484586 Fig. 4. Effects of FOXC2 silencing on invasion ability of HCC cell lines. (A) Representative images and quantification of the effects of FOXC2 silencing on the migratory and invasive abilities of MHCC-97H cells as determined by Transwell assays. (B) Representative images and quantification of the effects of FOXC2 silencing on the migratory and invasive abilities of Hep3B cells as determined by Transwell assays. (C) Western blot analysis showing downregulation of BCL-2, cleaved PARP, cleaved Caspase-3, p-AKT, MMP-2 and MMP-9 in FOXC2 and upregulation of cleaved PARP and cleaved Caspase-3 in FOXC2-depleted MHCC-97H and Hep3B cells. **P < 0.01. © 2017 The Author(s). Published by S. Karger AG, Basel - CC BY-NC-ND 4.0

Elevated FOXC2 Expression Promotes Invasion of HCC Cell Lines and is Associated with Poor Prognosis in Hepatocellular Carcinoma Cell Physiol Biochem 2017;44:99–109 - DOI:10.1159/000484586 Fig. 5. Knockdown of FOXC2 inhibits tumorigenesis of HCC cells. (A) Representative picture of tumor formation in the Hep3B scramble group or FOXC2-depleted Hep3B group in nude mice. (B) Tumor volumes were measured every week. (C) Tumor weight was decreased in the FOXC2-depleted Hep3B group mouse model. (D) Representation of FOXC2 and Ki-67 expression in xenograft tumors using IHC in the Hep3B scramble group or the FOXC2-depleted Hep3B group. **P < 0.01. © 2017 The Author(s). Published by S. Karger AG, Basel - CC BY-NC-ND 4.0