目的 研究重组反义c myc腺病毒 (Ad AS c myc)对人急性早幼粒HL 60细胞系的诱导分化作用及分子机制 ,探讨Ad AS c myc用于急性早幼粒白血病基因治疗的可能性。方法 采用重组腺病毒Lac Z(Ad LacZ)及Ad AS c myc联合多聚阳离子转染HL 6...目的 研究重组反义c myc腺病毒 (Ad AS c myc)对人急性早幼粒HL 60细胞系的诱导分化作用及分子机制 ,探讨Ad AS c myc用于急性早幼粒白血病基因治疗的可能性。方法 采用重组腺病毒Lac Z(Ad LacZ)及Ad AS c myc联合多聚阳离子转染HL 60细胞 ,X gal染色判断转染效率。采用形态学、MTT试验、RT PCR、流式细胞仪及免疫细胞化学等方法进行研究。结果 多聚阳离子可提高腺病毒对HL 60细胞的转染 ,Ad LacZ +protaminesulfate转染率达 79 8%。Ad AS c myc能抑制HL 60细胞c myc基因在转录及蛋白水平的表达。Ad AS c myc抑制HL 60细胞生长及活力 (抑制率5 1%) ,降低其核浆比例 ,增加其过氧化酶活性 ,引起其G0 G1 阻滞及分化相关基因c fos基因表达增强。结论 Ad AS c myc对HL 60细胞具有抑制生长及诱导分化作用。其生物学作用与HL 60细胞过氧化酶活性及c fos的表达有关。Ad AS c展开更多
This study investigated the effect of cadmium on the telomerase activity,the expression of TERT,c-myc and p53 and the apoptosis of rat hepatocytes.The rats were administrated 5,10 and 20 μmol/kg cadmium chloride intr...This study investigated the effect of cadmium on the telomerase activity,the expression of TERT,c-myc and p53 and the apoptosis of rat hepatocytes.The rats were administrated 5,10 and 20 μmol/kg cadmium chloride intraperitoneally and sacrificed 48 h after the initial treatment.The telomerase activity of the rat hepatocytes was measured by the telomeric repeat amplification protocol (TRAP),and apoptosis was detected by flow cytometry.The mRNA expressions of TERT,c-myc and p53 were measured by reverse transcription-polymerase chain reaction (RT-PCR).C-myc and P53 proteins were determined by immunochemistry.The results showed that cadmium chloride increased the hepatocellular telomerase activity in a dose-dependant manner and induced the apoptosis of hepatocytes significantly.The value of relative coefficient between the telomerase activity and the apoptosis rate was 0.9398.RT-PCR revealed that specific bands corresponding to the TERT mRNA,c-myc mRNA,and p53 mRNA were displayed at 185,342 and 538 bp respectively.Cadmium chloride could substantially increase the mRNA expressions of TERT,c-myc and p53 in rat hepatocytes,as compared with control.Moreover,cadmium chloride at the doses of 5,10 and 20 μmol/kg could increase the content of P53 protein in rat hepatocytes obviously,but only that at the doses of 10 and 20 μmol/kg substantially promoted the c-myc protein level in rat hepatocytes.Our study herein suggested that cadmium may contribute to the carcinogenesis by activating telomerase,and overexpressing the mRNAs of TERT,c-myc and p53,and causing apoptosis of normal cells.展开更多
文摘目的 研究重组反义c myc腺病毒 (Ad AS c myc)对人急性早幼粒HL 60细胞系的诱导分化作用及分子机制 ,探讨Ad AS c myc用于急性早幼粒白血病基因治疗的可能性。方法 采用重组腺病毒Lac Z(Ad LacZ)及Ad AS c myc联合多聚阳离子转染HL 60细胞 ,X gal染色判断转染效率。采用形态学、MTT试验、RT PCR、流式细胞仪及免疫细胞化学等方法进行研究。结果 多聚阳离子可提高腺病毒对HL 60细胞的转染 ,Ad LacZ +protaminesulfate转染率达 79 8%。Ad AS c myc能抑制HL 60细胞c myc基因在转录及蛋白水平的表达。Ad AS c myc抑制HL 60细胞生长及活力 (抑制率5 1%) ,降低其核浆比例 ,增加其过氧化酶活性 ,引起其G0 G1 阻滞及分化相关基因c fos基因表达增强。结论 Ad AS c myc对HL 60细胞具有抑制生长及诱导分化作用。其生物学作用与HL 60细胞过氧化酶活性及c fos的表达有关。Ad AS c
基金supported by the National Natural Sciences Foundation of China(No.30271110)Scientific Foundation(No.2006GGW01)Teacher's Foundation of Guangdong Pharmaceutical University,China
文摘This study investigated the effect of cadmium on the telomerase activity,the expression of TERT,c-myc and p53 and the apoptosis of rat hepatocytes.The rats were administrated 5,10 and 20 μmol/kg cadmium chloride intraperitoneally and sacrificed 48 h after the initial treatment.The telomerase activity of the rat hepatocytes was measured by the telomeric repeat amplification protocol (TRAP),and apoptosis was detected by flow cytometry.The mRNA expressions of TERT,c-myc and p53 were measured by reverse transcription-polymerase chain reaction (RT-PCR).C-myc and P53 proteins were determined by immunochemistry.The results showed that cadmium chloride increased the hepatocellular telomerase activity in a dose-dependant manner and induced the apoptosis of hepatocytes significantly.The value of relative coefficient between the telomerase activity and the apoptosis rate was 0.9398.RT-PCR revealed that specific bands corresponding to the TERT mRNA,c-myc mRNA,and p53 mRNA were displayed at 185,342 and 538 bp respectively.Cadmium chloride could substantially increase the mRNA expressions of TERT,c-myc and p53 in rat hepatocytes,as compared with control.Moreover,cadmium chloride at the doses of 5,10 and 20 μmol/kg could increase the content of P53 protein in rat hepatocytes obviously,but only that at the doses of 10 and 20 μmol/kg substantially promoted the c-myc protein level in rat hepatocytes.Our study herein suggested that cadmium may contribute to the carcinogenesis by activating telomerase,and overexpressing the mRNAs of TERT,c-myc and p53,and causing apoptosis of normal cells.
