NIH 3T3 cells, a mouse fibroblast cell line used as routine target cells for transfection experiments, undergo spontaneous transformation in our experiments after they form a confluent sheet in medium containing fetal...NIH 3T3 cells, a mouse fibroblast cell line used as routine target cells for transfection experiments, undergo spontaneous transformation in our experiments after they form a confluent sheet in medium containing fetal bovina serum (FBS) or lower coneentrafcion of calf serum (CS). The transformation takes the form of foci of multiplying cells among the surrounding cells which have stopped cell division. However, no focus of transformed cells could be seen in medium containing high concentration (10%) of OS. Further experiments indicated that the frequency of transformation is highly dependent on the concentration of serum and the transformation in OS is changeable when the cells are passaged in FBS. 8H-thymidine autoradiography has been proved to be a sensitive measurement indicator for foous formation. Our results suggest that the high frequency of transformation and its dependence on confmenoy as well as on medium composition are characteristics of cell differentiation rather than mutation. The role of the NIH 3T3 cell line as a cancer-initiated cell population and its accelerated transformation by rat oncogene might be considered as a form of tumor promotion is discussed.展开更多
For electronic microscopic observation, we found SSV-transformed NIH 3T3 cells were different from non-transformed cells. In SSV-transformed NIH 3T3 cells nuclei cytoplasma ratio was increased and in cytoplasma the ri...For electronic microscopic observation, we found SSV-transformed NIH 3T3 cells were different from non-transformed cells. In SSV-transformed NIH 3T3 cells nuclei cytoplasma ratio was increased and in cytoplasma the ribosomes (polyribosomes were attached to the swollen rough endoplasmic reticulum. It was likely that ribosomes were lined together functionally and structionally to produce specific protein (PDGF-like protein).展开更多
Lanthanum ions （La3＋） have been reported to exert profound effects on the proliferation of NIH 3T3 mouse embryonic fibroblasts. In the present studies, the pro-proliferative effect of La3＋ on NIH 3T3 cells was fur...Lanthanum ions （La3＋） have been reported to exert profound effects on the proliferation of NIH 3T3 mouse embryonic fibroblasts. In the present studies, the pro-proliferative effect of La3＋ on NIH 3T3 cells was further characterized, and its impacts on the cell cycle kinetics were analyzed in detail by cell cycle arrest-and-release and pulse BrdU-labeling and chasing experiments. The results show that La3＋ promoted the proliferation of NIH 3T3 cells after 12 h and the ECs0 was 2.4 ktM after 48 h. Such effect was also confirmed by enhanced BrdU incorporation. La3＋ stimulated more cells to pass through the G1/S checkpoint into S phase, but did not change the length of cell cycle. Furthermore, La3＋-treatment increased the protein levels and nuclear localization of cyclin D 1 and c-Myc, two key regulators of G1/S checkpoint, as demonstrated by immunostaining. The pro-proliferative effects of La3＋ share striking similarity with that of Gd3＋, and provided new information regarding the cellular biological activities of lanthanides.展开更多
Intracellular communications between breast cancer and fibroblast cells were reported to be involved in cancer proliferation,growth,and therapy resitance.The hallmarks of cancer fibroblast interactions,consisting of c...Intracellular communications between breast cancer and fibroblast cells were reported to be involved in cancer proliferation,growth,and therapy resitance.The hallmarks of cancer fibroblast interactions,consisting of caveolin 1(Cav1)and mono-carboxylate ransporter 4(MCT4)(metabolic coupling markers),along with IL-6,TGFB,and lactate secretion,are considered robust biomarkers predicting recurrence and metastasis.In order to promote a novel phenotype in normal fibroblasts,we predicted that breast cancer cells could be able to cause loss of Cavl and increase of MCT4,as well as elevate IL 6 and TGF in nearby nomal fibroblasts.We created a co culture model using breast cancer(4T1)and normal fibroblast(NIH3T3)cell lines cultured under specific experimental conditions in order to directly test our theory.Moreover,we show that long-term co-culture of breast cancer cells and normal fibroblasts promotes loss of Cavl and gain of MCT4 in adjacent fibroblasts and increase lactate secretion.These results were validated using the monoculture of each group separately as a control.In this system,we show that me tformin inhibits IL-6 and TGFB secretion and re expresses Cavl in both cells.However,MCT4 and lactate stayed high after treatment with metformin.In conclusion,our work shows that co-culture with breast cancer cells may cause signifcant alterations in the phenotype and secretion of normal fibroblasts.Metformin,however,may change this state and affect fibroblasts'acquired phenotypes.Moreover,mitochondrial inhibition by metformin after 8 days of treatment,signi ficantly hinders tumor growth in mouse model of breast cancer.展开更多
文摘NIH 3T3 cells, a mouse fibroblast cell line used as routine target cells for transfection experiments, undergo spontaneous transformation in our experiments after they form a confluent sheet in medium containing fetal bovina serum (FBS) or lower coneentrafcion of calf serum (CS). The transformation takes the form of foci of multiplying cells among the surrounding cells which have stopped cell division. However, no focus of transformed cells could be seen in medium containing high concentration (10%) of OS. Further experiments indicated that the frequency of transformation is highly dependent on the concentration of serum and the transformation in OS is changeable when the cells are passaged in FBS. 8H-thymidine autoradiography has been proved to be a sensitive measurement indicator for foous formation. Our results suggest that the high frequency of transformation and its dependence on confmenoy as well as on medium composition are characteristics of cell differentiation rather than mutation. The role of the NIH 3T3 cell line as a cancer-initiated cell population and its accelerated transformation by rat oncogene might be considered as a form of tumor promotion is discussed.
文摘For electronic microscopic observation, we found SSV-transformed NIH 3T3 cells were different from non-transformed cells. In SSV-transformed NIH 3T3 cells nuclei cytoplasma ratio was increased and in cytoplasma the ribosomes (polyribosomes were attached to the swollen rough endoplasmic reticulum. It was likely that ribosomes were lined together functionally and structionally to produce specific protein (PDGF-like protein).
基金National Natural Science Foundation (Grant No.21001011)Key Program of National Natural Science Foundation of China (Grant No. 41130746)
文摘Lanthanum ions （La3＋） have been reported to exert profound effects on the proliferation of NIH 3T3 mouse embryonic fibroblasts. In the present studies, the pro-proliferative effect of La3＋ on NIH 3T3 cells was further characterized, and its impacts on the cell cycle kinetics were analyzed in detail by cell cycle arrest-and-release and pulse BrdU-labeling and chasing experiments. The results show that La3＋ promoted the proliferation of NIH 3T3 cells after 12 h and the ECs0 was 2.4 ktM after 48 h. Such effect was also confirmed by enhanced BrdU incorporation. La3＋ stimulated more cells to pass through the G1/S checkpoint into S phase, but did not change the length of cell cycle. Furthermore, La3＋-treatment increased the protein levels and nuclear localization of cyclin D 1 and c-Myc, two key regulators of G1/S checkpoint, as demonstrated by immunostaining. The pro-proliferative effects of La3＋ share striking similarity with that of Gd3＋, and provided new information regarding the cellular biological activities of lanthanides.
基金the National Institute for Medical Research Development(NIMADGrant No.995813).
文摘Intracellular communications between breast cancer and fibroblast cells were reported to be involved in cancer proliferation,growth,and therapy resitance.The hallmarks of cancer fibroblast interactions,consisting of caveolin 1(Cav1)and mono-carboxylate ransporter 4(MCT4)(metabolic coupling markers),along with IL-6,TGFB,and lactate secretion,are considered robust biomarkers predicting recurrence and metastasis.In order to promote a novel phenotype in normal fibroblasts,we predicted that breast cancer cells could be able to cause loss of Cavl and increase of MCT4,as well as elevate IL 6 and TGF in nearby nomal fibroblasts.We created a co culture model using breast cancer(4T1)and normal fibroblast(NIH3T3)cell lines cultured under specific experimental conditions in order to directly test our theory.Moreover,we show that long-term co-culture of breast cancer cells and normal fibroblasts promotes loss of Cavl and gain of MCT4 in adjacent fibroblasts and increase lactate secretion.These results were validated using the monoculture of each group separately as a control.In this system,we show that me tformin inhibits IL-6 and TGFB secretion and re expresses Cavl in both cells.However,MCT4 and lactate stayed high after treatment with metformin.In conclusion,our work shows that co-culture with breast cancer cells may cause signifcant alterations in the phenotype and secretion of normal fibroblasts.Metformin,however,may change this state and affect fibroblasts'acquired phenotypes.Moreover,mitochondrial inhibition by metformin after 8 days of treatment,signi ficantly hinders tumor growth in mouse model of breast cancer.
