Exp Mol Med.  2009 Feb;41(2):102-115. 10.3858/emm.2009.41.2.013.

Genetic and expression alterations in association with the sarcomatous change of cholangiocarcinoma cells

• Yoo HJ
• Yun BR
• Kwon JH
• Ahn HS
• Seol MA
• Lee MJ
• Yu GR
• Yu HC
• Hong B
• Choi K
• Kim DG

• KMID: 1109433
• DOI: http://doi.org/10.3858/emm.2009.41.2.013

Abstract

Cholangiocarcinoma (CC) is an intrahepatic bile duct carcinoma with a high mortality rate and a poor prognosis. Sarcomatous change/epithelial mesenchymal transition (EMT) of CC frequently leads to aggressive intrahepatic spread and metastasis. The aim of this study was to identify the genetic alterations and gene expression pattern that might be associated with the sarcomatous change in CC. Previously, we established 4 human CC cell lines (SCK, JCK1, Cho-CK, and Choi-CK). In the present study, we characterized a typical sarcomatoid phenotype of SCK, and classified the other cell lines according to tumor cell differentiation (a poorly differentiated JCK, a moderately differentiated Cho-CK, and a well differentiated Choi-CK cells), both morphologically and immunocytologically. We further analyzed the genetic alterations of two tumor suppressor genes (p53 and FHIT) and the expression of Fas/FasL gene, well known CC-related receptor and its ligand, in these four CC cell lines. The deletion mutation of p53 was found in the sarcomatoid SCK cells. These cells expressed much less Fas/FasL mRNAs than did the other ordinary CC cells. We further characterize the gene expression pattern that is involved in the sarcomatous progression of CC, using cDNA microarrays that contained 18,688 genes. Comparison of the expression patterns between the sarcomatoid SCK cells and the differentiated Choi-CK cells enabled us to identify 260 genes and 247 genes that were significantly over-expressed and under-expressed, respectively. Northern blotting of the 14 randomly selected genes verified the microarray data, including the differential expressions of the LGALS1, TGFBI, CES1, LDHB, UCHL1, ASPH, VDAC1, VIL2, CCND2, S100P, CALB1, MAL2, GPX1, and ANXA8 mRNAs. Immunohistochemistry also revealed in part the differential expressions of these gene proteins. These results suggest that those genetic and gene expression alterations may be relevant to the sarcomatous change/EMT in CC cells.

Keyword

cholangiocarcinoma; gene expression profiling; oligonucleotide array sequence analysis; sarcoma

MeSH Terms

Acid Anhydride Hydrolases/genetics
Animals
Cell Line, Tumor
Cholangiocarcinoma/*genetics
Female
*Gene Expression Profiling
Humans
Mice
Mice, Inbred BALB C
Mutation
Neoplasm Proteins/genetics
Oligonucleotide Array Sequence Analysis
Sarcoma/*genetics
Tumor Suppressor Protein p53/genetics