Isolation of a human gallbladder cancer cell clone with high invasive phenotype in vitro and metastatic potential in orthotopic model and inhibition of its invasiveness by heparanase antisense oligodeoxynucleotides.
The mechanisms involved in gallbladder cancer metastasis still remain unclear to date. The poor understanding is due, in part, to the lack of ideal cell line and animal model for study. In the present study, 21 cell clones were isolated from the human gallbladder carcinoma cells GBC-SD and the cell clone GBC-SDH(i) with high invasive phenotype was fished out. The invasive phenotype and metastatic potential of GBC-SDH(i) were confirmed in a novel surgical orthotopic implantation model of gallbladder cancer in nude mice. Heparanase, an endoglycosidase that degrades heparan sulfate, is a critical mediator of tumor metastasis and angiogenesis. RT-PCR, real time RT-PCR and western blot showed that the expression levels of heparanase were significant difference between GBC-SDH(i) and its parent cells. After treated with antisense oligodeoxynucleotides, the heparanase mRNA and protein expression in GBC-SDH(i) cells were significantly decreased and its invasive potential in vitro was inhibited in a dose-dependent manner. The study provides a useful cell clone and a clinically relevant orthotopic tumor model for the metastatic study in human gallbladder cancer. The roles of heparanase in gallbladder cancer are also evaluated.
Chang
Xin-Zhong
XZ
Central Laboratory, Cancer Hospital/Cancer Institute, Fudan University, Shanghai 200032, P. R. China.
Wang
Zhan-Min
ZM
Yu
Jin-Ming
JM
Tian
Fu-Guo
FG
Jin
Wei
W
Zhang
Yi
Y
Yu
Jie
J
Li
Lian-Fang
LF
Liu
Xiao-Feng
XF
Li
Zhi-Wei
ZW
Shao
Zhi-Min
ZM
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002999:Clone Cells; D004195:Disease Models, Animal; D024202:Electrophoretic Mobility Shift Assay; D005706:Gallbladder Neoplasms / Q000201:enzymology* / Q000473:pathology; D005966:Glucuronidase / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D016376:Oligonucleotides, Antisense; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099175:Heparanase
D005966:Glucuronidase; D016376:Oligonucleotides, Antisense; D000099175:Heparanase