摘要
目的 探讨CML28分子在急性髓系白血病（acute myeloid leukemia, AML）细胞系K562细胞增殖与凋亡调控中的作用机制。方法 体外培养K562细胞，电转染特异性CML28小分子干扰RNA（small interfering RNA, siRNA），收集转染特异性Si-hCML2848h的K562细胞，Trizol一步法提取细胞总RNA，逆转录合成cDNA，通过反转录RCR (reverse transcription PCR, RT-PCR）及实时定量PCR（real time PCR）检测其沉默效应；流式细胞术(flow cytometry ,FCM)和CFSE标记的细胞增殖试验检测干扰后K562细胞的凋亡与增殖情况，SPSS17.0软件进行统计学分析，分类资料采用非参数检验，计量资料采用方差分析或t检验，以P <0.05为有统计学意义。 结果 （1）K562细胞高表达CML28分子；（2）干扰后CML28分子的mRNA水平为0.524 ±0.044，较阴性对照组明显下降, 非参数检验Kruskal-Wallis H：P=0.003, 检验水准α=0.05, P<α,差异有统计学意义；（3）Si-hCML28干扰组凋亡率为(22.45±3.54)%，较阴性对照组显著增高，方差分析：P=0.005,检验水准α=0.05, P<α,差异有统计学意义；Si-hCML28干扰后K562细胞增殖显著受抑，t检验：P=0.01, 检验水准α=0.05，P<α，差异有统计学意义。 结论 （1）CML28分子在K562细胞中呈高表达，Si-hCML28可以显著抑制CML28分子的mRNA表达；（2）siRNA干扰CML28分子后，显著抑制K562细胞的增殖，诱导K562细胞的凋亡。这些结果提示CML28在K562细胞的增殖与凋亡调控中扮演着重要的角色。
Abstract
Objective To explore the role of CML28 in regulation of proliferation and apoptosis of K562 cells. Methods K562 cells were cultured in vitro and electrotransfected with Si-hCML28. K562 cells which had been transfected were collected after 48 hours. Total mRNA was extracted from K562 cells using Trizol reagent and the first cDNA was synthesized using the reverse transcription kit.The silence effect was detected by RT-PCR and Real time PCR. The ratio of apoptosis and proliferation of K562 cells was detected by Flow Cytometry (FCM)after CML28 was down-regulated. Data were analyzed by SPSS17.0 statistics software and P <0.05 was considered statistically significant. Results (1) There was a high level of CML28 in K562 cells; (2)The mRNA expression of CML28 was 0.524 ± 0.044 after interference, lower than that of negative control. There was statistically significant difference；(3) The apoptosis ratio was (22.45±3.54)% in the group of Si-hCML28, higher than that of negative control. Conclusions CML28 is highly expressed in K562 cells. The mRNA expression of CML28 can be effectively down-regulated by Si-hCML28. In addition, it can significantly inhibit K562 cell proliferation and induce apoptosis of K562 cells, suggesting that it plays an important role in regulating the apoptosis and proliferation of K562 cells.
关键词
CML28 /
EXOSOME /
凋亡 /
增殖 /
急性髓系白血病
白雪玲,毛 霞,张 冰,曹丰斌,张东华.
CML28在K562细胞增殖与凋亡调控中的作用研究[J]. 武警医学. 2012, 23(8): 678-678
BAI Xueling,MAO Xia,ZHANG Bing,CAO Fengbin,and ZHANG Donghua..
The role of CML28 in regulation of proliferation and apoptosis of K562 cells[J]. Medical Journal of the Chinese People Armed Police Forces. 2012, 23(8): 678-678
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参考文献
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