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Establishment and characterisation of six human biliary tract cancer cell lines.

Ku JL, Yoon KA, Kim IJ, Kim WH, Jang JY, Suh KS, Kim SW, Park YH, Hwang JH, Yoon YB, Park JG - Br. J. Cancer (2002)

Bottom Line: In addition, we compared the genetic alterations in tumour cell lines and their corresponding tumour tissues.The culture success rate was 20% (six out of 30 attempts).Among the lines, three lines had p53 mutations; and homozygous deletions in both p16 and p15 genes were found three and three lines, respectively; one line had a heterozygous missense mutation in hMLH1; E-cadherin gene was hypermethylated in two lines.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell Biology, Korean Cell Line Bank, Cancer Research Center and Cancer Research Institute, Seoul National University College of Medicine, Seoul 110-744, Korea.

ABSTRACT
Human cell lines established from biliary tract cancers are rare, and only five have been reported previously. We report the characterisation of six new six biliary tract cancer cell lines (designated SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079 and SNU-1196) established from primary tumour samples of Korean patients. The cell lines were isolated from two extrahepatic bile duct cancers (one adenocarcinoma of common bile duct, one hilar bile duct cancer), two adenocarcinomas of ampulla of Vater, one intrahepatic bile duct cancer (cholangiocarcinoma), and one adenocarcinoma of the gall bladder. The cell phenotypes, including the histopathology of the primary tumours and in vitro growth characteristics, were determined. We also performed molecular characterisation, including DNA fingerprinting analysis and abnormalities of K-ras, p15, p16, p53, hMLH1, hMSH2, DPC4, beta-catenin, E-cadherin, hOGG1, STK11, and TGF-betaRII genes by PCR-SSCP and sequencing analysis. In addition, we compared the genetic alterations in tumour cell lines and their corresponding tumour tissues. All lines grew as adherent cells. Population doubling times varied from 48-72 h. The culture success rate was 20% (six out of 30 attempts). All cell lines showed (i) relatively high viability; (ii) absence of mycoplasma or bacteria contamination; and (iii) genetic heterogeneity by DNA fingerprinting analysis. Among the lines, three lines had p53 mutations; and homozygous deletions in both p16 and p15 genes were found three and three lines, respectively; one line had a heterozygous missense mutation in hMLH1; E-cadherin gene was hypermethylated in two lines. Since the establishment of biliary tract cancer cell lines has been rarely reported in the literature, these newly established and well characterised biliary tract cancer cell lines would be very useful for studying the biology of biliary tract cancers, particularly those related to hypermethylation of E-cadherin gene in biliary tract cancer.

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PCR-based deletion analysis of the p15 and p16 genes. (A,B) PCR amplification of the p15 gene. (C) PCR amplification of exon 2 in the K-ras gene used as internal control for separate PCR amplification. (D,E) PCR amplification of the p16 gene. (F) K-ras gene for internal control for separate PCR amplification. Lane numbers 1 to 9 show cell lines SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079, SNU-1196, HeLa, K-562, and water only.
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fig4: PCR-based deletion analysis of the p15 and p16 genes. (A,B) PCR amplification of the p15 gene. (C) PCR amplification of exon 2 in the K-ras gene used as internal control for separate PCR amplification. (D,E) PCR amplification of the p16 gene. (F) K-ras gene for internal control for separate PCR amplification. Lane numbers 1 to 9 show cell lines SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079, SNU-1196, HeLa, K-562, and water only.

Mentions: There were no band shifts in p15 and p16 genes by PCR-PCR SSCP analysis. However, there were no PCR amplifications in three lines (SNU-478, SNU-1079, and SNU-1196). By based deletion analysis of each gene, we found that exon 1 of p15 gene was not amplified in three cell lines (SNU-478, SNU-1079 and SNU-1196) and exon 2 of this gene was not amplified in two cell lines (SNU-478 and SNU-119). We also found that exons 1 and 2 of p16 gene were not amplified in three cell lines (SNU-478, SNU-1079 and SNU-1196) (Figure 4Figure 4


Establishment and characterisation of six human biliary tract cancer cell lines.

Ku JL, Yoon KA, Kim IJ, Kim WH, Jang JY, Suh KS, Kim SW, Park YH, Hwang JH, Yoon YB, Park JG - Br. J. Cancer (2002)

PCR-based deletion analysis of the p15 and p16 genes. (A,B) PCR amplification of the p15 gene. (C) PCR amplification of exon 2 in the K-ras gene used as internal control for separate PCR amplification. (D,E) PCR amplification of the p16 gene. (F) K-ras gene for internal control for separate PCR amplification. Lane numbers 1 to 9 show cell lines SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079, SNU-1196, HeLa, K-562, and water only.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376107&req=5

fig4: PCR-based deletion analysis of the p15 and p16 genes. (A,B) PCR amplification of the p15 gene. (C) PCR amplification of exon 2 in the K-ras gene used as internal control for separate PCR amplification. (D,E) PCR amplification of the p16 gene. (F) K-ras gene for internal control for separate PCR amplification. Lane numbers 1 to 9 show cell lines SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079, SNU-1196, HeLa, K-562, and water only.
Mentions: There were no band shifts in p15 and p16 genes by PCR-PCR SSCP analysis. However, there were no PCR amplifications in three lines (SNU-478, SNU-1079, and SNU-1196). By based deletion analysis of each gene, we found that exon 1 of p15 gene was not amplified in three cell lines (SNU-478, SNU-1079 and SNU-1196) and exon 2 of this gene was not amplified in two cell lines (SNU-478 and SNU-119). We also found that exons 1 and 2 of p16 gene were not amplified in three cell lines (SNU-478, SNU-1079 and SNU-1196) (Figure 4Figure 4

Bottom Line: In addition, we compared the genetic alterations in tumour cell lines and their corresponding tumour tissues.The culture success rate was 20% (six out of 30 attempts).Among the lines, three lines had p53 mutations; and homozygous deletions in both p16 and p15 genes were found three and three lines, respectively; one line had a heterozygous missense mutation in hMLH1; E-cadherin gene was hypermethylated in two lines.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell Biology, Korean Cell Line Bank, Cancer Research Center and Cancer Research Institute, Seoul National University College of Medicine, Seoul 110-744, Korea.

ABSTRACT
Human cell lines established from biliary tract cancers are rare, and only five have been reported previously. We report the characterisation of six new six biliary tract cancer cell lines (designated SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079 and SNU-1196) established from primary tumour samples of Korean patients. The cell lines were isolated from two extrahepatic bile duct cancers (one adenocarcinoma of common bile duct, one hilar bile duct cancer), two adenocarcinomas of ampulla of Vater, one intrahepatic bile duct cancer (cholangiocarcinoma), and one adenocarcinoma of the gall bladder. The cell phenotypes, including the histopathology of the primary tumours and in vitro growth characteristics, were determined. We also performed molecular characterisation, including DNA fingerprinting analysis and abnormalities of K-ras, p15, p16, p53, hMLH1, hMSH2, DPC4, beta-catenin, E-cadherin, hOGG1, STK11, and TGF-betaRII genes by PCR-SSCP and sequencing analysis. In addition, we compared the genetic alterations in tumour cell lines and their corresponding tumour tissues. All lines grew as adherent cells. Population doubling times varied from 48-72 h. The culture success rate was 20% (six out of 30 attempts). All cell lines showed (i) relatively high viability; (ii) absence of mycoplasma or bacteria contamination; and (iii) genetic heterogeneity by DNA fingerprinting analysis. Among the lines, three lines had p53 mutations; and homozygous deletions in both p16 and p15 genes were found three and three lines, respectively; one line had a heterozygous missense mutation in hMLH1; E-cadherin gene was hypermethylated in two lines. Since the establishment of biliary tract cancer cell lines has been rarely reported in the literature, these newly established and well characterised biliary tract cancer cell lines would be very useful for studying the biology of biliary tract cancers, particularly those related to hypermethylation of E-cadherin gene in biliary tract cancer.

Show MeSH
Related in: MedlinePlus