Anat Cell Biol.  2010 Jun;43(2):110-117. 10.5115/acb.2010.43.2.110.

CDH3/P-Cadherin regulates migration of HuCCT1 cholangiocarcinoma cells

• Baek S
• Lee YW
• Yoon S
• Baek SY
• Kim BS
• Oh SO

• KMID: 2168883
• DOI: http://doi.org/10.5115/acb.2010.43.2.110

Abstract

Intrahepatic cholangiocarcinoma is the second most common subtype of primary hepatobilliary cancer. Despite advances in surgical and medical therapy, its survival rate remains poor. Compared to hepatocellular carcinoma (HCC), the most common liver malignancy, the underlying mechanisms of cholangiocarcinoma carcinogenesis are poorly characterized. P-cadherin (CDH3) is a cadherin super family member. Although CDH3 is frequently over-expressed in cholangiocarcinoma tissues, its roles have never been characterized. To determine the roles of CDH3 in cholangiocarcinoma, we investigated CDH3 function in HuCCT1 cells using specific siRNA. Transfection with CDH3 siRNA did not affect proliferation of HuCCT1 cells. However, cell migration and invasion were significantly reduced when CDH3 was down-regulated. In addition, expressions of several biomarkers for epithelial-mesenchymal transition (EMT) were not changed by CDH3 down-regulation. These results suggest that CDH3 regulates cell migration independent of EMT in cholangiocarcinoma cells.

Keyword

Cholangiocarcinoma; CDH3; migration

MeSH Terms

Biomarkers
Cadherins
Carcinoma, Hepatocellular
Cell Movement
Cholangiocarcinoma
Down-Regulation
Epithelial-Mesenchymal Transition
Humans
Liver
Liver Neoplasms
RNA, Small Interfering
Survival Rate
Transfection
Cholangiocarcinoma
Cadherins
Cholangiocarcinoma
Liver Neoplasms
RNA, Small Interfering

Figure

• 

  Fig. 1 CDH3 siRNA specifically knock-downed CDH3 expression level. 48 h following transfection, CDH3 and SCR siRNA-transfected HuCCT1 cells were collected and RNA and protein were purified. Real-time RT-PCR (A) and Western blotting (B) were performed with CDH3 specific primers and antibody. β-actin was used for loading control. Data are expressed as percent change (means±SEM) compared to control. *P<0.01 (Student's t-test).

• 

  Fig. 2 Cell proliferation is not affected by down-regulation of CDH3 in HuCCT1 cells. SCR siRNA and different concentration series (100 nM, 50 nM, 10 nM, and 1 nM) of CDH3 siRNA-transfected HuCCT1 cells in a 96-well plate were incubated for 4 d, and MTT assay was performed. Data are expressed as percent change (means±SEM) compared to control.

• 

  Fig. 3 Down-regulation of CDH3 in HuCCT1 cells reduced migration in wound healing assay. After transfection as described in Materials and Methods, HuCCT1 cells were scratched and allowed to migrate up to 22 h. Control = no transfected, SCR = Scrambled siRNA transfected.

• 

  Fig. 4 Down-regulation of CDH3 in HuCCT1 cells reduced invasion in Matrigel invasion assay. After transfection as described in Materials and Methods, HuCCT1 cells were allowed to invade through the matrigel up to 36 h. Invaded cells were stained with hematoxylin/eosin (A) and counted to quantify (B). Data are expressed as percent change (means±SEM) compared to control. *P<0.01 (Student's t-test).

• 

  Fig. 5 Expression of EMT-related genes in CDH3-knockdowned HuCCT1 cells. RNA was purified from SCR or CDH3 siRNA-transfected cells and real-time RT-PCR was carried out. After normalization to β-actin, data are expressed as percent change (means±SEM) compared to control.

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