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目的研究体外扩增过程中,CD34+细胞内B细胞特异单克隆鼠白血病毒整合位点基因1(B-cell-specific monoclonal leukemia virus insert site 1,Bmi1)和人端粒酶逆转录酶(human telomerase reverse transcriptase,hTERT)基因的表达水平与其扩增特性的相关性。方法采用含FBS、干细胞生长因子、Flt-3配体和促血小板生成素的IMDM培养基体外培养脐血CD34+细胞,在28 d培养过程中检测CD34+细胞扩增倍数、CD34+细胞比生长速率以及集落形成率的变化趋势,并采用荧光定量PCR检测体外扩增过程中CD34+细胞内Bmi1和hTERT基因表达水平变化,分析以上基因表达水平与CD34+细胞扩增特性之间的关系。结果经过28 d的体外培养,CD34+细胞共扩增了(20.1±3.5)倍,其占扩增后总细胞的比例由培养前的95.5%±2.6%下降至2.1%±0.4%;CD34+细胞的比生长速率和集落形成率均在培养7 d后出现明显下降,而细胞内Bmi1和hTERT mRNA的表达水平在培养7 d时达最高,此后逐渐下降至培养前水平。结论 Bmi1和hTERT基因表达与CD34+细胞体外增殖能力可能存在一定相关性。
OBJECTIVE: To investigate the role of Bcl-1 in Bcl-2 expression in human CD34 + cells during the in vitro expansion of human lung adenocarcinoma reverse transcriptase, hTERT) gene expression and its amplification characteristics. Methods Cord blood CD34 + cells were cultured in vitro with IMDM medium containing FBS, stem cell growth factor, Flt-3 ligand and thrombopoietin. The multiplication rate of CD34 + cells, specific growth rate of CD34 + cells, The changes of the expression of Bmi1 and hTERT in CD34 + cells were detected by fluorescence quantitative PCR. The relationship between the above gene expression and the amplification characteristics of CD34 + cells was analyzed. Results After 28 days of culture, the total number of CD34 + cells expanded (20.1 ± 3.5) times and the percentage of total cells after expansion decreased from 95.5% ± 2.6% to 2.1% ± 0.4% The specific growth rate and colony formation rate decreased significantly after 7 days of culture. However, the expression of Bmi1 and hTERT mRNA reached the peak at 7 days after culture, and then gradually decreased to pre-culture level. Conclusion The expression of Bmi1 and hTERT may be related to the proliferation of CD34 + cells in vitro.