Diverse cellular transformation capability of overexpressed genes in human hepatocellular carcinoma.
For isolation of novel cellular transforming genes that potentially participated in hepatocarcinogenesis, we conducted anchorage-independent growth (AIG) assays on 10 human liver cancer cell lines and observed strong AIG capabilities in PLC5 and Huh7 but negligible in Tong cells. After cloning of genes by differential subtractive chain reactions (DSC) from strong AIG to AIG negative cells, we sequenced 2304 clones and identified 245 genes. After four stringent criteria for selection of transforming genes among DSC clones, our results of quantitative RT-PCR analysis indicated that six genes, DDX3, EIF3S2, CLIC1, HDGF, GPC3, and HSPCA were overexpressed in 64%, 62%, 60%, 58%, 49%, and 47%, respectively, of 45 hepatocellular carcinoma (HCC) tissues. The results of cellular transformation capability by AIG assays indicated that the transfectants of EIF3S2 showed the strongest (> 100-fold), DDX3 and CLIC1 were moderate, GPC3 and HSPCA were weak, and HDGF was none in forming colonies in soft agar. Together, our results suggested that Tong is a suitable human cell line for screening of overexpressed and/or cellular transforming genes. In addition, our results suggested that diverse functions of cellular transforming genes in various biological pathways could transform human Tong cells and potentially reveal new targets for drug development of HCC.
Huang
Jhy-Shrian
JS
Graduate Institute of Life Sciences, National Defense Medical Center, National Defense University, Taipei, Taiwan, ROC.
Chao
Chuan-Chuan
CC
Su
Teh-Li
TL
Yeh
Shiou-Hwei
SH
Chen
Ding-Shinn
DS
Chen
Chiung-Tong
CT
Chen
Pei-Jer
PJ
Jou
Yuh-Shan
YS
D015152:Blotting, Northern / Q000379:methods; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003001:Cloning, Molecular; D003584:Cytological Techniques / Q000379:methods; D020869:Gene Expression Profiling; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D009857:Oncogenes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D014162:Transfection
D009363:Neoplasm Proteins