Oytoskeletal changes in transformed cells (LM-51) exhibiting obviously metastatie eapabilities were investigated by utilization of double-fluorescent labelling through combinations of:(1) tubulin indirect immunofluore...Oytoskeletal changes in transformed cells (LM-51) exhibiting obviously metastatie eapabilities were investigated by utilization of double-fluorescent labelling through combinations of:(1) tubulin indirect immunofluoresoenoe plus Khodamine-phalloidin staining of F-artins;(2) indirect immunofluorescent staining with α-aotinin polyolonal- and vinoulin monoclonal antibodies. The LM-51 cells which showed metastatic index of >50% were derived from lung metastasis in nude mice after subcutaneous inoculation of human highly metastatic tumor DNA transfected NIH3T3 cell transformants. The parent NIH3T3 cells exhibited well-organized miorotubu-les, prominent stress fibers and adhesion plaques while their transformants showed remarkable oytoskeletal alterations: (1) reduced microtubules but increased MTOC fluorescence; (2) disrupted stress fibers and fewer adhesion plaques with their protein components redistributed in the cytoplasm; (3) F-aotin-and α-actinin/vinculin aggregates appeared in the cytoplasm. These aggregates were dot-like, varied in size (0.1-0.4u,m) and number, located near the ventral surface of the cells. TPA-induced aotin/vinoulin bodies were studied too. Indications that aotin and α-actinin/vinoulin redistribution might be important alterations involved in the expression of metastatio capabilities of LM-51 transformed cells were discussed.展开更多
文摘Oytoskeletal changes in transformed cells (LM-51) exhibiting obviously metastatie eapabilities were investigated by utilization of double-fluorescent labelling through combinations of:(1) tubulin indirect immunofluoresoenoe plus Khodamine-phalloidin staining of F-artins;(2) indirect immunofluorescent staining with α-aotinin polyolonal- and vinoulin monoclonal antibodies. The LM-51 cells which showed metastatic index of >50% were derived from lung metastasis in nude mice after subcutaneous inoculation of human highly metastatic tumor DNA transfected NIH3T3 cell transformants. The parent NIH3T3 cells exhibited well-organized miorotubu-les, prominent stress fibers and adhesion plaques while their transformants showed remarkable oytoskeletal alterations: (1) reduced microtubules but increased MTOC fluorescence; (2) disrupted stress fibers and fewer adhesion plaques with their protein components redistributed in the cytoplasm; (3) F-aotin-and α-actinin/vinculin aggregates appeared in the cytoplasm. These aggregates were dot-like, varied in size (0.1-0.4u,m) and number, located near the ventral surface of the cells. TPA-induced aotin/vinoulin bodies were studied too. Indications that aotin and α-actinin/vinoulin redistribution might be important alterations involved in the expression of metastatio capabilities of LM-51 transformed cells were discussed.
