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背景与目的阿糖胞苷(Ara-C)是治疗急性非淋巴细胞白血病常用而有效的药物之一。端粒酶是一种特殊的核糖核蛋白复合物,在肿瘤的发生、发展中起着重要作用。本研究以Ara-C为诱导剂,检测了不同浓度、不同时间Ara-C作用HL-60细胞端粒酶各组分基因mRNA水平的变化,试图为临床阿糖胞苷的用药和评判化疗药效提供一些理论上的依据和指导。方法采用不同浓度Ara-C作用相同时间、同一浓度Ara-C作用不同时间来处理HL-60细胞,通过流式细胞仪观察其凋亡和坏死细胞比例,RT-PCR方法检测端粒酶各组分基因hTERT、hTR和hTP1的mRNA水平变化。结果①体外小剂量0~0.2μg/ml的Ara-C诱导HL-60细胞12h,端粒酶各组分基因在mRNA水平上无变化;②2μg/ml和10μg/mlAra-C组,端粒酶hTERTmRNA水平由0.80±0.07分别下调至0.50±0.04和0.39±0.03而hTR、hTP1mRNA无变化;③10μg/ml浓度的Ara-C作用HL-60细胞不同时间,随时间推移,细胞凋亡比例增加,12h处达到最大值(18.16±4.25)%,随后降低;而细胞坏死比率一直随时间的延长而增加,48h处的坏死比例在本实验的时间段处最大,为(57.94±12.03)%,同时hTERT基因mRNA水平也随着时间的延长而逐渐降低,48h处达最低值0.18±0.03,而hTR、hTP1mRNA水平不随时间的延长而改变。结论①阿糖胞苷下调hTERT基因的mRNA水平,且有浓度和时间依赖性,而对hTR基因、hTP1基因的mRNA水平无影响;②阿糖胞苷下调hTERT基因mRNA水平可能与阿糖胞苷诱导细胞坏死有关,而与诱导细胞凋亡无关。
Background and objective Cytarabine (Ara-C) is one of the commonly used and effective drugs for the treatment of acute non-lymphocytic leukemia. Telomerase is a kind of special ribonucleoprotein complex, which plays an important role in tumorigenesis and development. In this study, Ara-C was used as an inducer to detect the mRNA levels of telomerase mRNA in HL-60 cells treated with Ara-C at different concentrations and different times. The aim of this study was to evaluate the clinical effects of cytarabine and chemotherapeutic drugs Effective to provide some theoretical basis and guidance. Methods HL-60 cells were treated with different concentrations of Ara-C for the same time, with the same concentration of Ara-C for different time. The percentage of apoptosis and necrosis cells was detected by flow cytometry and the expression of telomerase was detected by RT- The mRNA levels of hTERT, hTR and hTP1 genes were analyzed. Results ① The HL-60 cells were treated with 0 ~ 0.2μg / ml Ara-C in a small dose for 12h, the mRNA level of telomerase did not change. ② The mRNA levels of telomerase The hTERT mRNA levels were down-regulated from 0.80 ± 0.07 to 0.50 ± 0.04 and 0.39 ± 0.03, respectively, while no change was found in hTR1 mRNA; ③ Ara-C at a concentration of 10μg / ml increased the percentage of apoptotic cells in HL-60 cells over time, (18.16 ± 4.25)%, and then decreased. However, the rate of cell necrosis increased with the increase of time. The proportion of necrosis at 48h was the largest (57.94 ± 12.03)% at the time of this experiment, while hTERT The gene mRNA level also decreased gradually with time, reaching the lowest value of 0.18 ± 0.03 at 48h, while hTR, hTP1mRNA level did not change with time. Conclusion ① Cytosine down-regulated the mRNA level of hTERT gene in a concentration- and time-dependent manner, but had no effect on the mRNA levels of hTR gene and hTP1 gene. ② Cytosine down-regulated the mRNA level of hTERT gene and cytarabine Induction of cell necrosis, but not apoptosis induction.