TY - JOUR
T1 - Suppression of in vitro invasion and in vivo growth of breast cancer induced by hepatocyte growth factor/scatter factor (HGF/SF) and HGF/SF producing fibroblasts, by viral hammerhead ribozymes targeting HGF/SF and its receptor cMET
AU - Jiang, W. G.
AU - Grimshaw, D. A.
AU - Lane, J.
AU - Martin, T. A.
AU - Laterra, J.
AU - Mansel, R. E.
PY - 2001/1/1
Y1 - 2001/1/1
N2 - Hepatocyte growth factor/scatter factor (HGF/SF) is known to increase the migration and invasiveness of breast cancer cells. This study aimed to target the impact of HGF/SF on the growth of breast cancer, by using hammerhead ribozymes. Methods. Retroviral hammerhead ribozymes were constructed to specifically target human HGF/SF and its receptor cMET. A breast cancer cell, MDA MB 231, which expressed cMET was transduced with viral cMET ribozyme. A human fibroblast cell, MRC5, that produces bioactive HGF/SF was transduced with HGF/SF ribozyme. In vitro invasiveness of both wild and transduced breast cancer cells were analysed using a Matrigel invasion assay. Production of HGF/SF from wild and transduced fibroblasts was analysed using MDCK bioassay. Breast cancer cells (wild and stable transducants) were used in a nude mice model, to determine the growth of breast tumours. Results. MET ribozyme eliminated the expression of cMET mRNA, as shown by RT-PCR. The stably transduced MDA MB 231 cells lost its response to HGF/SF in the in vitro invasion assay (invasion index being 2.3±0.9 for wild, vs 0.99±0.18 in the transduced, p<0.05). HGF/SF ribozyme reduced production of bioactive HGF/SF from fibroblasts (62±31U/ml from wild vs 3.4±1.6 U/ml from the transduced, p<0.01). As a consequence, the transduced fibroblasts reduced their effect on the invasiveness on breast cancer cells, in a co-culture invasion assay (invasion index 3.1 ± 1 with wild fibroblasts vs 1.4±0.6 with transduced fibroblasts, p<0.01). In in vivo studies, MET-transduced breast cancer cells exhibited a significant reduction in growth (tumour volume 1.84±1.6mm3 with wild and 0.26±0.2mm3 with tranduced tumour cells over 4 wks period, p=0.05). Conclusion. Viral hammerhead ribozymes are effective in targeting HGF/SF receptor and HGF/SF production by fibroblasts. These ribozymes are therefore useful in suppressing HGF/SF induced invasion and tumour growth of breast cancer and may have potential therapeutic value.
AB - Hepatocyte growth factor/scatter factor (HGF/SF) is known to increase the migration and invasiveness of breast cancer cells. This study aimed to target the impact of HGF/SF on the growth of breast cancer, by using hammerhead ribozymes. Methods. Retroviral hammerhead ribozymes were constructed to specifically target human HGF/SF and its receptor cMET. A breast cancer cell, MDA MB 231, which expressed cMET was transduced with viral cMET ribozyme. A human fibroblast cell, MRC5, that produces bioactive HGF/SF was transduced with HGF/SF ribozyme. In vitro invasiveness of both wild and transduced breast cancer cells were analysed using a Matrigel invasion assay. Production of HGF/SF from wild and transduced fibroblasts was analysed using MDCK bioassay. Breast cancer cells (wild and stable transducants) were used in a nude mice model, to determine the growth of breast tumours. Results. MET ribozyme eliminated the expression of cMET mRNA, as shown by RT-PCR. The stably transduced MDA MB 231 cells lost its response to HGF/SF in the in vitro invasion assay (invasion index being 2.3±0.9 for wild, vs 0.99±0.18 in the transduced, p<0.05). HGF/SF ribozyme reduced production of bioactive HGF/SF from fibroblasts (62±31U/ml from wild vs 3.4±1.6 U/ml from the transduced, p<0.01). As a consequence, the transduced fibroblasts reduced their effect on the invasiveness on breast cancer cells, in a co-culture invasion assay (invasion index 3.1 ± 1 with wild fibroblasts vs 1.4±0.6 with transduced fibroblasts, p<0.01). In in vivo studies, MET-transduced breast cancer cells exhibited a significant reduction in growth (tumour volume 1.84±1.6mm3 with wild and 0.26±0.2mm3 with tranduced tumour cells over 4 wks period, p=0.05). Conclusion. Viral hammerhead ribozymes are effective in targeting HGF/SF receptor and HGF/SF production by fibroblasts. These ribozymes are therefore useful in suppressing HGF/SF induced invasion and tumour growth of breast cancer and may have potential therapeutic value.
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M3 - Article
AN - SCOPUS:33749104921
VL - 69
JO - Breast Cancer Research and Treatment
JF - Breast Cancer Research and Treatment
SN - 0167-6806
IS - 3
ER -