To investigate the modulating effects of survivn antisense oligonucletode (ASODN) on the cell cycle and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721 and explore its mechanism.Methods Survivin ...To investigate the modulating effects of survivn antisense oligonucletode (ASODN) on the cell cycle and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721 and explore its mechanism.Methods Survivin ASODN was transfected into SMMC-7721 cells mediated by DOTAP liposomal reagent.Electron microscopy,flow cytometry and RT-PCR were used to detect the changes in cell ultrastructure,apoptosis,cell cycle and the expression of cyclinB1 mRNA,respectively.Results After transfection of survivin ASODN,the expression of cyclinB1 mRNA in the cells significantly increased and increase in G2-M arrest and apoptosis appeared.Meanwhile,the cell ultrastructure had apoptotic changes such as chromatin condensation and apoptotic body formation.Conclusion Survivin ASODN can induce the expression of cyclinB1 that may result in G2-M arrest.Consequently,apoptosis is triggered.Survivin ASODN transfection might be an improtant new treatment for HCC.14 refs,2 figs,1 tab.展开更多
Artemisinin, also termed qinghaosu, is extracted from the traditional Chinese medicine ar- temesia annua L. （the blue-green herb） in the early 1970s, which has been confirmed for effectively treating malaria, Additi...Artemisinin, also termed qinghaosu, is extracted from the traditional Chinese medicine ar- temesia annua L. （the blue-green herb） in the early 1970s, which has been confirmed for effectively treating malaria, Additionally, emerging data prove that artemisinin exhibits anti-cancer effects against many types of cancers such as leukemia, melanoma, etc. Artemisinin becomes cytotoxic in the presence of ferrous iron. Since iron influx is high in cancer cells, artemisinin and its analogs selectively kill can- cer cells with increased intracellular iron concentrations. This study is aimed to investigate the selective inhibitory effects of artemisinin on SMMC-7721 cells in vitro and determine the effect of holotransfer- fin, which increases the concentration of ferrous iron in cancer cells, combined with artemisinin on the anticancer activity. MTT assay was used for assessing the proliferation of SMMC-7721 cells treated with artemisinin. The induction of apoptosis and inhibition of colony formation in SMMC-7721 cells treated with artemisinin were determined by TdT-mediated dUTP nick end labeling （TUNEL） and col- ony formation assay, respectively. The results showed that artemisinin at various concentrations signifi- cantly inhibited growth, colony formation and cell viability of SMMC-7721 cells （P〈0.05）, likely due to induction of apoptosis of SMMC-7721 cells. Of interest, it was found that incubation of artemisinin combined with holotransferrin sensitized the growth inhibitory effect of artemisinin on SMMC-7721 cells （P〈0.01）. Our data suggest that treatment with artemisinin leads to inhibition of viability and pro- liferation, and apoptosis of SMMC-7721 ceils. Furthermore, we observed that holotransferrin signifi- cantly enhanced the anti-cancer activity of artemisinin. This study may provide a potential therapeutic choice for liver cancer.展开更多
AIM: To determine the radiosensitizing potential of docetaxel in human hepatocellular carcinoma SMMC-7721 cells and its mechanisms.METHODS: SMMC-7721 cells were incubated with docetaxel at 0.125, 0.25, and 0.5 nmoL/L ...AIM: To determine the radiosensitizing potential of docetaxel in human hepatocellular carcinoma SMMC-7721 cells and its mechanisms.METHODS: SMMC-7721 cells were incubated with docetaxel at 0.125, 0.25, and 0.5 nmoL/L for 24 h and at 0.125 and 0.25 nmol/L for 48 h before irradiation. Radiation doses were given from 0 to 10 Gy. Cell survival was measured by a standard clonogenic assay after a 9-d incubation. The reactive oxygen species (ROS) and glutathione (GSH) are detected after being given the same dose of docetaxel for the same time. RESULTS: The sensitization enhancement ratios (SER) for SMMC-7721 cells determined at the 50% survival level were 1.15, 1.21 and 1.49 at 0.125, 0.25, and 0.5 nmol/L for pre-incubation of 24 h, respectively; the SER were 1.42, 1.67 at 0.125 and 0.25 nmol/L, for pre-incubation of 48 h, respectively. The ROS of SMMC-7721 cells increased and GSH decreased after pretreatment with the same doses of docetaxel for 24 or 48 h.CONCLUSION: A radiosensitizing effect of docetaxel could be demonstrated unambiguously in this cell line used. In addition, our data showed that the mechanism of radiopotentiation by docetaxel probably does not involve a G2/M block in SMMC-7721 cells, and ROS generation and GSH deletion may play a key role in the radiosensitizing effect of docetaxel.展开更多
AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells ...AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells in vitro and on mouse implanted S-180tumors in vivo.METHODS The polysaccharide extracts were isolated from the fruit bodies of FV, LE and AB by the methods of hot-water extraction, Sevag’sremoval of proteins, ethanol precipitation,trypsin digestion and ethanol fractionalprecipitation. Human hepatoma SMMC-7721 cells were treated with 50 mg/L Polysaccharide extracts, and the mitosis index, mitochondria activity and cell proliferation were detected at different times in both control and experimental groups. The mice with S-180 implanted tumors were injected with the polysaccharide extracts at 24 mg/ kg body weight for 9 d and the tumorweight was measured on the 15th day.RESULTS The mitosis index of hepatoma cells in vitro could be significantly decreased by treatment with the polysaccharide extracts fromthe three kinds of edible fungi (P < 0 .005 ). Thecell numbers and mitochondria activity of SMMC7721 cells treated with polysaccharide extracts were lower than those in control groups (P <0.005). The inhibition rates of polysaccharide extracts against implanted S-180 tumors in mice were 52.8%, 56.6% and 51 .9% respectivelycompared with that in c0ntrol gr0ups.CONCLUSI0N The POIysaccharide extractsfrom the three kinds of edible fungi could inhibitnot only the Cultured malignant cells in vitfO butalso impIanted Sl80 tum0r i0 vivo.展开更多
AIM To determine the relationship between serum deprivation or serum levels and cell proliferation of human hepatoma SMMC 7721 cells. METHODS Human hepatoma SMMC 7721 cells were grown in RPMI 1640 supplemented wi...AIM To determine the relationship between serum deprivation or serum levels and cell proliferation of human hepatoma SMMC 7721 cells. METHODS Human hepatoma SMMC 7721 cells were grown in RPMI 1640 supplemented with 10% fetal calf (FCS) in 5% CO 2 incubator at 37℃ for 24h , and culture media were replaced to serum free or different serum FCS levels (2 5%, 5%, 10%, 20% and 25%). Six h, 12h , 18h and 24h after the culture, the cells were incorporated TdR for 4h . At last TdR incorporation was detected with liquid scintillation counting. RESULTS DNA synthesis of SMMC 7721 cells could be sharply stimulated by short time (6h) serum deprivation (the cpm value of 3H TdR incorporation of cells in serum free was 39 32 fold higher than cells in 25% serum), and the incorporation of 3H TdR was negatively related to the serum levels. Longer time serum starvation ( 12h , 18h and 24h ) also greatly stimulated DNA synthesis, although the cpm value of 3H TdR incroporation was less than that in 6h serum deprivation. Morphology of cells cultured in different serum levels also showed significant difference. CONCLUSIONS Compared with other cell lines such as BEL7404 and Swiss 3T3, human hepatoma SMMC 7721 cells had different response to the serum deprivation. Short time serum deprivation could greatly stimulate DNA synthesis of human hepatoma SMMC 7721 cells. Precautions must be given to the changes of serum levels for the detection of growth factors and drugs using SMMC 7721 cells as a model.展开更多
文摘To investigate the modulating effects of survivn antisense oligonucletode (ASODN) on the cell cycle and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721 and explore its mechanism.Methods Survivin ASODN was transfected into SMMC-7721 cells mediated by DOTAP liposomal reagent.Electron microscopy,flow cytometry and RT-PCR were used to detect the changes in cell ultrastructure,apoptosis,cell cycle and the expression of cyclinB1 mRNA,respectively.Results After transfection of survivin ASODN,the expression of cyclinB1 mRNA in the cells significantly increased and increase in G2-M arrest and apoptosis appeared.Meanwhile,the cell ultrastructure had apoptotic changes such as chromatin condensation and apoptotic body formation.Conclusion Survivin ASODN can induce the expression of cyclinB1 that may result in G2-M arrest.Consequently,apoptosis is triggered.Survivin ASODN transfection might be an improtant new treatment for HCC.14 refs,2 figs,1 tab.
文摘Artemisinin, also termed qinghaosu, is extracted from the traditional Chinese medicine ar- temesia annua L. （the blue-green herb） in the early 1970s, which has been confirmed for effectively treating malaria, Additionally, emerging data prove that artemisinin exhibits anti-cancer effects against many types of cancers such as leukemia, melanoma, etc. Artemisinin becomes cytotoxic in the presence of ferrous iron. Since iron influx is high in cancer cells, artemisinin and its analogs selectively kill can- cer cells with increased intracellular iron concentrations. This study is aimed to investigate the selective inhibitory effects of artemisinin on SMMC-7721 cells in vitro and determine the effect of holotransfer- fin, which increases the concentration of ferrous iron in cancer cells, combined with artemisinin on the anticancer activity. MTT assay was used for assessing the proliferation of SMMC-7721 cells treated with artemisinin. The induction of apoptosis and inhibition of colony formation in SMMC-7721 cells treated with artemisinin were determined by TdT-mediated dUTP nick end labeling （TUNEL） and col- ony formation assay, respectively. The results showed that artemisinin at various concentrations signifi- cantly inhibited growth, colony formation and cell viability of SMMC-7721 cells （P〈0.05）, likely due to induction of apoptosis of SMMC-7721 cells. Of interest, it was found that incubation of artemisinin combined with holotransferrin sensitized the growth inhibitory effect of artemisinin on SMMC-7721 cells （P〈0.01）. Our data suggest that treatment with artemisinin leads to inhibition of viability and pro- liferation, and apoptosis of SMMC-7721 ceils. Furthermore, we observed that holotransferrin signifi- cantly enhanced the anti-cancer activity of artemisinin. This study may provide a potential therapeutic choice for liver cancer.
文摘AIM: To determine the radiosensitizing potential of docetaxel in human hepatocellular carcinoma SMMC-7721 cells and its mechanisms.METHODS: SMMC-7721 cells were incubated with docetaxel at 0.125, 0.25, and 0.5 nmoL/L for 24 h and at 0.125 and 0.25 nmol/L for 48 h before irradiation. Radiation doses were given from 0 to 10 Gy. Cell survival was measured by a standard clonogenic assay after a 9-d incubation. The reactive oxygen species (ROS) and glutathione (GSH) are detected after being given the same dose of docetaxel for the same time. RESULTS: The sensitization enhancement ratios (SER) for SMMC-7721 cells determined at the 50% survival level were 1.15, 1.21 and 1.49 at 0.125, 0.25, and 0.5 nmol/L for pre-incubation of 24 h, respectively; the SER were 1.42, 1.67 at 0.125 and 0.25 nmol/L, for pre-incubation of 48 h, respectively. The ROS of SMMC-7721 cells increased and GSH decreased after pretreatment with the same doses of docetaxel for 24 or 48 h.CONCLUSION: A radiosensitizing effect of docetaxel could be demonstrated unambiguously in this cell line used. In addition, our data showed that the mechanism of radiopotentiation by docetaxel probably does not involve a G2/M block in SMMC-7721 cells, and ROS generation and GSH deletion may play a key role in the radiosensitizing effect of docetaxel.
文摘AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells in vitro and on mouse implanted S-180tumors in vivo.METHODS The polysaccharide extracts were isolated from the fruit bodies of FV, LE and AB by the methods of hot-water extraction, Sevag’sremoval of proteins, ethanol precipitation,trypsin digestion and ethanol fractionalprecipitation. Human hepatoma SMMC-7721 cells were treated with 50 mg/L Polysaccharide extracts, and the mitosis index, mitochondria activity and cell proliferation were detected at different times in both control and experimental groups. The mice with S-180 implanted tumors were injected with the polysaccharide extracts at 24 mg/ kg body weight for 9 d and the tumorweight was measured on the 15th day.RESULTS The mitosis index of hepatoma cells in vitro could be significantly decreased by treatment with the polysaccharide extracts fromthe three kinds of edible fungi (P < 0 .005 ). Thecell numbers and mitochondria activity of SMMC7721 cells treated with polysaccharide extracts were lower than those in control groups (P <0.005). The inhibition rates of polysaccharide extracts against implanted S-180 tumors in mice were 52.8%, 56.6% and 51 .9% respectivelycompared with that in c0ntrol gr0ups.CONCLUSI0N The POIysaccharide extractsfrom the three kinds of edible fungi could inhibitnot only the Cultured malignant cells in vitfO butalso impIanted Sl80 tum0r i0 vivo.
文摘AIM To determine the relationship between serum deprivation or serum levels and cell proliferation of human hepatoma SMMC 7721 cells. METHODS Human hepatoma SMMC 7721 cells were grown in RPMI 1640 supplemented with 10% fetal calf (FCS) in 5% CO 2 incubator at 37℃ for 24h , and culture media were replaced to serum free or different serum FCS levels (2 5%, 5%, 10%, 20% and 25%). Six h, 12h , 18h and 24h after the culture, the cells were incorporated TdR for 4h . At last TdR incorporation was detected with liquid scintillation counting. RESULTS DNA synthesis of SMMC 7721 cells could be sharply stimulated by short time (6h) serum deprivation (the cpm value of 3H TdR incorporation of cells in serum free was 39 32 fold higher than cells in 25% serum), and the incorporation of 3H TdR was negatively related to the serum levels. Longer time serum starvation ( 12h , 18h and 24h ) also greatly stimulated DNA synthesis, although the cpm value of 3H TdR incroporation was less than that in 6h serum deprivation. Morphology of cells cultured in different serum levels also showed significant difference. CONCLUSIONS Compared with other cell lines such as BEL7404 and Swiss 3T3, human hepatoma SMMC 7721 cells had different response to the serum deprivation. Short time serum deprivation could greatly stimulate DNA synthesis of human hepatoma SMMC 7721 cells. Precautions must be given to the changes of serum levels for the detection of growth factors and drugs using SMMC 7721 cells as a model.
