title: Characterization of two novel cell lines with distinct heterogeneity derived from a single human bile duct carcinoma.
abstract: BACKGROUND
  Intratumoral heterogeneity reflects subclonal diversity and accounts for a variety of clinically defined phenotypes including the development of drug resistance and recurrence. However, intratumoral heterogeneity of bile duct carcinoma (BDC) is rarely studied.
  
  
  METHODS
  Two highly heterogeneous cell lines named EH-CA1a and EH-CA1b were established from a primary tumor tissue of a pathologically proven BDC. Distinct heterogeneity and underlying mechanisms of two cell lines in karyotype, colony formation, tumorgenicity, and sensitivity to chemoradiotherapy were intensively studied.
  
  
  RESULTS
  Both cell lines showed typical morphology of cancer cells. EH-CA1a cells grew as free-floating aggregates, while EH-CA1b cells grew adherently as a monolayer. EH-CA1a cells had higher cloning efficiencies and were able to keep proliferating under hypoxic condition. Coincidentally, hypoxia-induced factor-1α (HIF1α) and vascular endothelial growth factor (VEGF) mRNA were significantly higher in EH-CA1a cells than in EH-CA1b cells. Both cell lines were tumorigenic in nude mouse, however, EH-CA1a cells showed more aggressive characteristics. Most importantly, the EH-CA1a cells showed much more resistance against radiation and chemotherapy with gemcitabine. Metastasis-related genes including matrix metalloproteinase 2 (MMP-2), MMP-9, epithelial-mesenchymal transition (EMT) markers such as Vimentin, Snail, and Twist, are more highly expressed in EH-CA1a cells than in EH-CA1b cells. Moreover, the percentage of cells expressing cancer stem cell-like marker, CD133, in EH-CA1a cells is much higher than that in EH-CA1b cells. Moreover, knockdown of CD133 in both EH-CA1a and EH-CA1b cells significantly reduced their invasive potential and increased their sensitivities to radiation and gemcitabine, suggesting the differential expression of CD133 protein may partially account for the difference in malignancy between these two cancer cells.
  
  
  CONCLUSIONS
  Establishment of these two cell lines will not only shed light on intratumoral heterogeneities of BDC, but also potentially facilitate the development of novel therapeutic approaches of BDC.
authors: Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgical Hospital, The Second Military Medical University, Shanghai, China.
authors: Jinghan
authors: J
authors: Wang
authors: Linfang
authors: L
authors: Li
authors: Keqiang
authors: K
authors: Zhang
authors: Yong
authors: Y
authors: Yu
authors: Bin
authors: B
authors: Li
authors: Jiang
authors: J
authors: Li
authors: Zi
authors: Z
authors: Yan
authors: Zhenli
authors: Z
authors: Hu
authors: Yun
authors: Y
authors: Yen
authors: Mengchao
authors: M
authors: Wu
authors: Xiaoqing
authors: X
authors: Jiang
authors: Qijun
authors: Q
authors: Qian
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001650:Bile Duct Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D015415:Biomarkers; D002277:Carcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects / Q000528:radiation effects; D002869:Chromosome Aberrations; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D058750:Epithelial-Mesenchymal Transition; D055785:Gene Knockdown Techniques; D006801:Humans; D019656:Loss of Heterozygosity; D008297:Male; D051379:Mice; D053842:Microsatellite Instability; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000378:metabolism; D014183:Transplantation, Heterologous; D014410:Tumor Stem Cell Assay
chemical_list: D000970:Antineoplastic Agents; D015415:Biomarkers
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