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
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
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。
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.
