To investigate the synergistic effects of 3′-azido-3′- deoxythymidine （AZT） and FA-2-b-β extracted from Ling Jin mushroom on apoptosis of gastric cancer cells MKN45 in vitro. METHODS： Ml-I- analysis was made to ...To investigate the synergistic effects of 3′-azido-3′- deoxythymidine （AZT） and FA-2-b-β extracted from Ling Jin mushroom on apoptosis of gastric cancer cells MKN45 in vitro. METHODS： Ml-I- analysis was made to examine the inhibition rate of MKN45 cells treated with AZT （2.5, 5, 10 and 20 mg/L） and FA-2-b-13 （5, 10, 20 and 40 mg/L） singly and combinatively for 24, 48 and 72 h. Apoptotic effects were evaluated by morphological methods, DNA agarose gel electrophoresis and flow cytometry, respectively. Telomerase activity was estimated by TRAP- ELISA. The mRNA expression of caspase-3 and Bcl-2 were detected by RT-PCR. RESULTS： AZT and FA-2-b-13 could significantly inhibit MKN45 cell proliferation and induce its apoptosis. MKN45 cells were inhibited in dose- and time- dependent manner. The inhibition effect of AZT combined with FA-2- b-β was obviously better than that used singly （0.469 ＋ 0.022 vs 1.075 4- 0.055, P 〈 0.05, 0.325 4- 0.029 vs 0.469 ＋ 0.022 P 〈 0.01）. AZT used singly and combination of FA-2-b-β could decrease the activity of tumor cell telomerase, and AZT has synergistic function with FA- 2-b-β. A certain concentration of AZT could up-regulate the expression of caspase-3 mRNA （r = 0.9969, P 〈 0.01）, which was positively related to apoptosis rate, and could down-regulate the expression of Bcl-2 mRNA, which was negatively related to apoptosis rate （r = 0.926, P 〈 0.01）. Furthermore, the effect of AZT combined with FA-2-b-13 was significantly higher than that used singly. CONCLUSION： Combination of AZT and FA-2-b-β has an obviously synergetic effect in the gastric cancer cells MKN45, which has provided a new approach to the treatment of gastric cancer clinically.展开更多
A convenient, efficient and general method has been developed for synthesis of H-phosphonate mono and diesters of AZT and d4T through one-pot reaction of phosphonic acid with AZT or d4T and different alcohols using pi...A convenient, efficient and general method has been developed for synthesis of H-phosphonate mono and diesters of AZT and d4T through one-pot reaction of phosphonic acid with AZT or d4T and different alcohols using pivaloyl chloride as condensing agent under mild conditions.展开更多
近几年的研究显示通过抑端粒酶活性,干预端粒酶的激活过程,阻断肿瘤细胞的亚性转化,在肿瘤治疗上有重要意义和巨大的潜力[1]。本研究通过端粒酶抑制剂3-叠氮脱氧胸苷（3-azido-deoxythymidine,AZT）作用于人胃癌细胞SGC7901,比较AZT作...近几年的研究显示通过抑端粒酶活性,干预端粒酶的激活过程,阻断肿瘤细胞的亚性转化,在肿瘤治疗上有重要意义和巨大的潜力[1]。本研究通过端粒酶抑制剂3-叠氮脱氧胸苷（3-azido-deoxythymidine,AZT）作用于人胃癌细胞SGC7901,比较AZT作用后癌细胞端粒酶活性的变化,并利用缺口末端标记技术【terminal deoxyribonucleotidyl transferse（TdT）-mediated biotin-16dUTP nick end labeling,TUNEL】和流式细胞术检测AZT作用后肿瘤作用后胃癌细胞SGC7901凋亡情况,进而探讨AZT抑制肿瘤细胞端粒酶可能的机制,为端粒酶抑制剂的临床应用奠定基础。展开更多
文摘To investigate the synergistic effects of 3′-azido-3′- deoxythymidine （AZT） and FA-2-b-β extracted from Ling Jin mushroom on apoptosis of gastric cancer cells MKN45 in vitro. METHODS： Ml-I- analysis was made to examine the inhibition rate of MKN45 cells treated with AZT （2.5, 5, 10 and 20 mg/L） and FA-2-b-13 （5, 10, 20 and 40 mg/L） singly and combinatively for 24, 48 and 72 h. Apoptotic effects were evaluated by morphological methods, DNA agarose gel electrophoresis and flow cytometry, respectively. Telomerase activity was estimated by TRAP- ELISA. The mRNA expression of caspase-3 and Bcl-2 were detected by RT-PCR. RESULTS： AZT and FA-2-b-13 could significantly inhibit MKN45 cell proliferation and induce its apoptosis. MKN45 cells were inhibited in dose- and time- dependent manner. The inhibition effect of AZT combined with FA-2- b-β was obviously better than that used singly （0.469 ＋ 0.022 vs 1.075 4- 0.055, P 〈 0.05, 0.325 4- 0.029 vs 0.469 ＋ 0.022 P 〈 0.01）. AZT used singly and combination of FA-2-b-β could decrease the activity of tumor cell telomerase, and AZT has synergistic function with FA- 2-b-β. A certain concentration of AZT could up-regulate the expression of caspase-3 mRNA （r = 0.9969, P 〈 0.01）, which was positively related to apoptosis rate, and could down-regulate the expression of Bcl-2 mRNA, which was negatively related to apoptosis rate （r = 0.926, P 〈 0.01）. Furthermore, the effect of AZT combined with FA-2-b-13 was significantly higher than that used singly. CONCLUSION： Combination of AZT and FA-2-b-β has an obviously synergetic effect in the gastric cancer cells MKN45, which has provided a new approach to the treatment of gastric cancer clinically.
基金Project supported by Excellent Dissertation Foundation by the Chinese Ministry of Education (No. 200222).
文摘A convenient, efficient and general method has been developed for synthesis of H-phosphonate mono and diesters of AZT and d4T through one-pot reaction of phosphonic acid with AZT or d4T and different alcohols using pivaloyl chloride as condensing agent under mild conditions.
文摘近几年的研究显示通过抑端粒酶活性,干预端粒酶的激活过程,阻断肿瘤细胞的亚性转化,在肿瘤治疗上有重要意义和巨大的潜力[1]。本研究通过端粒酶抑制剂3-叠氮脱氧胸苷（3-azido-deoxythymidine,AZT）作用于人胃癌细胞SGC7901,比较AZT作用后癌细胞端粒酶活性的变化,并利用缺口末端标记技术【terminal deoxyribonucleotidyl transferse（TdT）-mediated biotin-16dUTP nick end labeling,TUNEL】和流式细胞术检测AZT作用后肿瘤作用后胃癌细胞SGC7901凋亡情况,进而探讨AZT抑制肿瘤细胞端粒酶可能的机制,为端粒酶抑制剂的临床应用奠定基础。
