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目的探讨骨髓增生异常综合征(MDS)患者骨髓细胞中P16基因表达异常态势,为寻找MDS患者分子诊断标记提供实验依据。方法采用荧光实时定量聚合酶联反应检测44例初诊和18例治疗后MDS患者骨髓单个核细胞中P16基因mRNA表达水平,并与15例非MDS贫血患者相比较。磁珠分选8例初诊MDS患者CD34+细胞,并用流式细胞仪检测细胞纯度。检测MDS患者骨髓单个核细胞及CD34+细胞中P16表达水平。结果在单个核细胞中,初诊MDS组与治疗后MDS组P16基因表达水平均显著高于非MDS贫血组,差异均有高度统计学意义(P<0.001)。流式细胞仪检测CD34+细胞纯度为96.7%,在CD34+细胞中,初诊MDS组P16基因表达水平显著高于非MDS贫血组,差异有统计学意义(P<0.05)。结论MDS患者P16表达异常可发生在CD34+细胞,也可发生在分化后较成熟的单个核细胞。P16基因可作为MDS鉴别诊断的分子标志物之一,其在MDS患者细胞凋亡中可能发挥作用。
Objective To investigate the abnormal expression of P16 gene in bone marrow cells of patients with myelodysplastic syndrome (MDS), and to provide experimental evidence for finding molecular diagnostic markers in patients with MDS. Methods The mRNA expression of P16 in bone marrow mononuclear cells from 44 newly diagnosed and 18 patients with MDS was detected by real-time fluorescence quantitative polymerase chain reaction and compared with 15 non-MDS anemia patients. The CD34 + cells from 8 newly diagnosed MDS patients were sorted by magnetic beads, and the cell purity was detected by flow cytometry. The level of P16 in bone marrow mononuclear cells and CD34 + cells of MDS patients was detected. Results In mononuclear cells, the expression of P16 gene in newly diagnosed MDS group and post-treatment MDS group were significantly higher than those in non-MDS anemia group, the differences were highly statistically significant (P <0.001). The purity of CD34 + cells detected by flow cytometry was 96.7%. The expression level of P16 gene in newly diagnosed MDS group was significantly higher than that in non-MDS anemia group in CD34 + cells (P <0.05). Conclusion The abnormal expression of P16 in MDS patients may occur in CD34 + cells and may also occur in mature mononuclear cells after differentiation. P16 gene can be used as a molecular marker for the differential diagnosis of MDS, which may play a role in the apoptosis of MDS patients.