Reduction of stromal fibroblast-induced mammary tumor growth, by retroviral ribozyme transgenes to hepatocyte growth factor/scatter factor and its receptor, c-MET.
OBJECTIVE
Hepatocyte growth factor/scatter factor (HGF/SF) is known to increase the invasiveness and migration of cancer cells in vitro and induce angiogenesis. This study examined if inhibition of HGF/SF receptor expression by cancer cells and HGF/SF expression by stromal fibroblasts affects the growth of mammary cancer.


METHODS
Transgenes encoding ribozymes to specifically target human HGF/SF receptor (pLXSN-MET) or HGF/SF (pLXSN-HGF) were constructed using a pLXSN retroviral vector. Human mammary cancer cells MDA MB 231 was transduced with pLXSN-MET (MDA(+/+)). A human fibroblast cell line MRC5, which produces bioactive HGF/SF, was transduced with pLXSN-HGF (MRC5(+/+)). These cells were used in a nude mice breast tumor model.


RESULTS
HGF receptor in MDA(+/+) cells and HGF in MRC5(+/+)cells were successfully removed with respective ribozymes as shown by reverse transcription-PCR and Western blotting, respectively. MDA(+/+) was found to have reduced invasiveness when stimulated with HGF/SF. MRC5(+/+) exhibited a significant reduction in HGF/SF production. When injected into athymic nude mice, MDA(+/+) exhibited a slower rate of growth, compared with the wild type (MDA(-/-)), and the cells transduced with control viral vector (MDA(+/-)). The growth of MDA(-/-) tumor was significantly enhanced when coimplanted with wild-type MRC5 (MRC5(-/-)), and the stimulatory effect was reduced when MRC5(+/+) cells were coimplanted instead of MRC5(-/-). The reduction of tumor growth was accompanied by reduction of angiogenesis, as demonstrated by the staining of VE-cadherin in primary tumor tissues.


CONCLUSIONS
Retroviral ribozyme transgenes targeting HGF/SF in fibroblasts or its receptor cMET in mammary cancer cells can reduce the growth of mammary cancer and associated angiogenesis by inhibiting paracrine stromal-tumor cell interactions.
D000818:Animals; D001943:Breast Neoplasms / Q000098:blood supply / Q000473:pathology*; D002455:Cell Division; D002460:Cell Line; D045744:Cell Line, Tumor; D005260:Female; D005347:Fibroblasts / Q000166:cytology*; D005822:Genetic Vectors; D017228:Hepatocyte Growth Factor / Q000235:genetics*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009389:Neovascularization, Pathologic / Q000517:prevention & control; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics*; D016337:RNA, Catalytic / Q000235:genetics; D015183:Restriction Mapping; D012190:Retroviridae; D017154:Stromal Cells / Q000166:cytology* / Q000473:pathology; D014162:Transfection; D014183:Transplantation, Heterologous
D016337:RNA, Catalytic; C427204:hammerhead ribozyme; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met