title: Screening candidate metastasis-associated genes in three-dimensional HCC spheroids with different metastasis potential.
abstract: OBJECTIVE
  Previously, we have established a tissue-like HCC spheroid which better mirrors the biological features of tumorigenesis and metastasis. This study was to find out metastasis-associated genes between two 3D HCC spheroids with different metastasis potential using comparative PCR arrays.
  
  
  METHODS
  Two HCC spheroids derived from high-metastatic MHCC97H cells and low-metastatic Hep3B cells were formed respectively in a rotating wall vessel bioreactor after 3D culture for 15 days. The candidate metastasis-associated genes related to cell adhesion, matrix secretion and invasion in HCC spheroids were screened by RT² profiler PCR arrays. The expression patterns of several differentially-expressed genes were further confirmed by real-time RT-PCR.
  
  
  RESULTS
  Total of 123 differential expression genes (fold-change>2) were found between two HCC spheroids, including 70 up-regulated genes (VCAM-1, IL-1β, CD44, tenascin C, SPP1, fibronectin, MMP-2, MMP-7, etc) and 53 down-regulated genes (E-cadherin, CTNND2, etc) in the high-metastatic spheroid. Function classification showed that the number of up-regulated genes related to adhesion molecules mediating cell-matrix interactions and matrix secretion was significantly higher in high-metastatic spheroid than that in low-metastatic spheroid. In contrast, the expressions of adhesion molecules maintaining homotypic tumor cell adhesion were decreased in metastatic spheroid as compared with that in low-metastatic spheroid. In addition, the expression pattern of seven selected genes associated with tumor metastasis measured by real-time RT-PCR were consistent with results of PCR arrays.
  
  
  CONCLUSIONS
  Obvious differences between two HCC spheroids in gene expression patterns of adhesion molecules, matrix secretion, invasion and other molecules may determine the different metastatic characteristics and malignant phenotype of HCC spheroid.
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Rongxin
authors: R
authors: Chen
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Yinying
authors: Y
authors: Dong
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Xiaoying
authors: X
authors: Xie
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Jie
authors: J
authors: Chen
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Dongmei
authors: D
authors: Gao
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Yinkun
authors: Y
authors: Liu
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Zhenggang
authors: Z
authors: Ren
authors: Liver Cancer Institute, Zhongshan Hospital, Fudan University; Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education 136 Xue Yuan Road, Shanghai 200032, China.
authors: Jiefeng
authors: J
authors: Cui
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics*; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000556:secondary*; D015815:Cell Adhesion Molecules / Q000235:genetics; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D016326:Extracellular Matrix Proteins / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic; D005838:Genotype; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000473:pathology*; D009361:Neoplasm Invasiveness; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D060888:Real-Time Polymerase Chain Reaction; D018874:Spheroids, Cellular
chemical_list: D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins
keywords: Hepatocellular carcinoma; metastasis; spheroid; three-dimensional culture