Objective To study the inhibitory effects of caspase-3 mRNA antisense oligodeoxynucleotides (ASODN) on expressions of caspase-3 and it's mRNA in γ-radiation induced apoptotic HL-60 cells, and screen the effective...Objective To study the inhibitory effects of caspase-3 mRNA antisense oligodeoxynucleotides (ASODN) on expressions of caspase-3 and it's mRNA in γ-radiation induced apoptotic HL-60 cells, and screen the effective ASODN. Methods ASODN-1 and ASODN-2 targeting 5′-noncoding region and initial translation region of caspase-3 mRNA were respectively designed, synthesized and introduced into HL-60 cells by means of liposome-mediated transfection followed by 10Gy γ-radiation exposures. TUNEL assay was conducted to investigate the morphologic change and apoptotic percentage of HL-60 cells 18 h later. Immunocytochemical staining and one step RT-PCR were respectively performed to detect the expressions of caspase-3 and it's mRNA. Mismatched oligodeoxynucleotide (MODN) transfected and un-transfected HL-60 cells were taken as control. Results TUNEL assay found that the apoptotic percentages in ASODN-1 and ASODN-2 groups were significantly reduced compared with the control groups (P<0.01) when the final concentration of both ASODNs was ≥3μmol/L. Immunocytochemistry showed that caspase-3 positive cell percentages were reduced but the average gray values increased significantly compared with the control groups (P<0.01). RT-PCR showed expressions of caspase-3 mRNA was decreased after ASODN transfection. Furthermore, ASODN-1 proved more effective in inhibiting HL-60 cell apoptosis than ASODN-2 (P<0.01). Conclusion Caspase-3 mRNA ASODNs can prevent HL-60 cells from apoptosis induced by γ-radiation and reduce expression of caspase-3 and its mRNA. These effects are dose dependent in a certain range.展开更多
Anticancer activity in vitro ofLitsea cubeba fruit extracts was investigated, focusing on the fruit residue from essential oil extraction. The methanol extract was fractionated by an Amberlite XAD-7 column. Cell viabi...Anticancer activity in vitro ofLitsea cubeba fruit extracts was investigated, focusing on the fruit residue from essential oil extraction. The methanol extract was fractionated by an Amberlite XAD-7 column. Cell viability, cell proliferation and cell death were determined using conversion of WST-8, BrdU incorporation and measurement of released LDH, respectively. Activation of caspase-3/-7 was detected using Z-DEVD-R substrate and morphological characteristics of apoptotic cells were revealed by DAPI staining. It was found that 80-100% methanol fractions （RME-4B, -5A, -5B and -5C） were effective against HeLa cell viability and also promoted cell death. RME-SA and -5B were highly effective in suppressing DNA replication （IC50 4.89 and 3.26 g/mL at 48 h） and also in activation of caspase-3/-7 （9 and 17 times of untreated population at 12 h）. The presence of apoptotic bodies was clearly observed. The results of this study suggested that L. cubeba fruit residue has remarkable apoptosis induction potential for further use in cancer drug research and for waste management in the essential oil industry.展开更多
AIM：To investigate the effects of Terminalia arjuna （T. arjuna） extract on human hepatoma cell line （HepG2） and its possible role in induction of apoptosis.METHODS： Human hepatoma cells were treated with differe...AIM：To investigate the effects of Terminalia arjuna （T. arjuna） extract on human hepatoma cell line （HepG2） and its possible role in induction of apoptosis.METHODS： Human hepatoma cells were treated with different concentrations of ethanolic extract of T. arjuna and its cytotoxicity effect was measured by trypan blue exclusion method and lactate dehydrogenase leakage assay. Apoptosis was analyzed by light and fluorescence microscopic methods, and DNA fragmentation. The mechanism of apoptosis was studied with expression of p53 and caspase-3 proteins. Glutathione （GSH） content was also measured in HepG2 cells after T. arjuna treatment.RESULTS： T. arjuna inhibited the proliferation of HepG2 cells in a concentration-dependent manner. Apoptotic morphology was observed in HepG2 cells treated with T. arjuna at the concentrations of 60 and 100 mg/L. DNA fragmentation, accumulation of p53 and cleavage of procaspase-3 protein were observed in HepG2 cells after the treatment with T. arjuna. The depletion of GSH was observed in HepG2 cells treated with T. arjuna.CONCLUSION： T. arjuna induced cytotoxicity in HepG2 cells in vitro. Apoptosis of HepG2 cells may be due to the DNA damage and expression of apoptotic proteins. Depletion of GSH may be involved in the induction of apoptosis of HepG2 cells.展开更多
文摘Objective To study the inhibitory effects of caspase-3 mRNA antisense oligodeoxynucleotides (ASODN) on expressions of caspase-3 and it's mRNA in γ-radiation induced apoptotic HL-60 cells, and screen the effective ASODN. Methods ASODN-1 and ASODN-2 targeting 5′-noncoding region and initial translation region of caspase-3 mRNA were respectively designed, synthesized and introduced into HL-60 cells by means of liposome-mediated transfection followed by 10Gy γ-radiation exposures. TUNEL assay was conducted to investigate the morphologic change and apoptotic percentage of HL-60 cells 18 h later. Immunocytochemical staining and one step RT-PCR were respectively performed to detect the expressions of caspase-3 and it's mRNA. Mismatched oligodeoxynucleotide (MODN) transfected and un-transfected HL-60 cells were taken as control. Results TUNEL assay found that the apoptotic percentages in ASODN-1 and ASODN-2 groups were significantly reduced compared with the control groups (P<0.01) when the final concentration of both ASODNs was ≥3μmol/L. Immunocytochemistry showed that caspase-3 positive cell percentages were reduced but the average gray values increased significantly compared with the control groups (P<0.01). RT-PCR showed expressions of caspase-3 mRNA was decreased after ASODN transfection. Furthermore, ASODN-1 proved more effective in inhibiting HL-60 cell apoptosis than ASODN-2 (P<0.01). Conclusion Caspase-3 mRNA ASODNs can prevent HL-60 cells from apoptosis induced by γ-radiation and reduce expression of caspase-3 and its mRNA. These effects are dose dependent in a certain range.
文摘Anticancer activity in vitro ofLitsea cubeba fruit extracts was investigated, focusing on the fruit residue from essential oil extraction. The methanol extract was fractionated by an Amberlite XAD-7 column. Cell viability, cell proliferation and cell death were determined using conversion of WST-8, BrdU incorporation and measurement of released LDH, respectively. Activation of caspase-3/-7 was detected using Z-DEVD-R substrate and morphological characteristics of apoptotic cells were revealed by DAPI staining. It was found that 80-100% methanol fractions （RME-4B, -5A, -5B and -5C） were effective against HeLa cell viability and also promoted cell death. RME-SA and -5B were highly effective in suppressing DNA replication （IC50 4.89 and 3.26 g/mL at 48 h） and also in activation of caspase-3/-7 （9 and 17 times of untreated population at 12 h）. The presence of apoptotic bodies was clearly observed. The results of this study suggested that L. cubeba fruit residue has remarkable apoptosis induction potential for further use in cancer drug research and for waste management in the essential oil industry.
文摘AIM：To investigate the effects of Terminalia arjuna （T. arjuna） extract on human hepatoma cell line （HepG2） and its possible role in induction of apoptosis.METHODS： Human hepatoma cells were treated with different concentrations of ethanolic extract of T. arjuna and its cytotoxicity effect was measured by trypan blue exclusion method and lactate dehydrogenase leakage assay. Apoptosis was analyzed by light and fluorescence microscopic methods, and DNA fragmentation. The mechanism of apoptosis was studied with expression of p53 and caspase-3 proteins. Glutathione （GSH） content was also measured in HepG2 cells after T. arjuna treatment.RESULTS： T. arjuna inhibited the proliferation of HepG2 cells in a concentration-dependent manner. Apoptotic morphology was observed in HepG2 cells treated with T. arjuna at the concentrations of 60 and 100 mg/L. DNA fragmentation, accumulation of p53 and cleavage of procaspase-3 protein were observed in HepG2 cells after the treatment with T. arjuna. The depletion of GSH was observed in HepG2 cells treated with T. arjuna.CONCLUSION： T. arjuna induced cytotoxicity in HepG2 cells in vitro. Apoptosis of HepG2 cells may be due to the DNA damage and expression of apoptotic proteins. Depletion of GSH may be involved in the induction of apoptosis of HepG2 cells.
