[Study on autophagy in nucleated red blood cells in patients with myelodysplastic syndromes]

Zhonghua Xue Ye Xue Za Zhi. 2017 May 14;38(5):432-436. doi: 10.3760/cma.j.issn.0253-2727.2017.05.015.
[Article in Chinese]

Abstract

Objective: To investigate the change of autophagy level of bone marrow nucleated red blood cell (RBC) in patients with myelodysplastic syndromes (MDS) . Methods: Fifty-four MDS patients and thirty-three controls were enrolled in this study. The mitophagy were observed by transmission electron microscopy (TEM) . The level of autophagy-associated protein LC3B in GlycoA(+) nucleated RBC was measured by flow cytometry. The expressions of ULK1 and mTOR mRNA in GlycoA(+) nucleated RBC were measured by real-time PCR. The expression of the mitochondrial outer membrane protein TOM20 in GlycoA(+) nucleated RBC was detected by Western blot. Results: Autophagosomes or autolysosomes were scarcely observed by TEM in MDS patients. The expression of LC3B in GlycoA(+) nucleated RBC in high-risk MDS patients (0.22±0.12) was significantly lower than that in normal controls (0.43±0.22, P<0.001) , and lower than that in low-risk MDS patients (0.40±0.16, P=0.001) . The expression of AMPK [0.26 (0.60) ] in GlycoA(+) nucleated RBC in high-risk MDS patients was significantly lower than that in controls [1.00 (2.07) , P<0.017) . The expression of ULK1 mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [0.27 (3.31) ] was significantly lower than that in controls [1.07 (4.41) , P<0.017]. The level of mTOR mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [1.82 (3.74) ] was significantly higher than that in controls [1.26 (1.38) , P<0.017]. The level of LC3B in GlycoA(+) nucleated RBC was negatively correlated with the HGB (r=0.529, P=0.009) in high-risk MDS patients. The expression of mitochondrial outer membrane protein TOM20 in high-risk MDS patients was 9.42±4.42. Conclusion: Autophagy is impaired in nucleated RBC of MDS patients.

目的: 研究骨髓增生异常综合征(MDS)患者有核红细胞的自噬水平。 方法: 54例MDS患者为实验组,对照组33例。采用透射电镜观察自噬过程,流式细胞术检测骨髓细胞中GlycoA(+)有核红细胞自噬相关蛋白微管相关蛋白轻链3B(LC3B)的表达水平,Real-time PCR法检测GlycoA(+)有核红细胞自噬相关基因AMPK、ULK1和mTOR mRNA的表达,Western blot法检测GlycoA(+)有核红细胞线粒体外膜蛋白TOM20的表达。 结果: MDS患者有核红细胞中不易观察到自噬体及自噬溶酶体。相对高危组MDS患者有核红细胞LC3B表达(0.22±0.12)显著低于对照组(0.43±0.22,P<0.001)及相对低危组(0.40±0.16,P=0.001)。相对高危组患者AMPK、ULK1 mRNA表达水平明显低于对照组[AMPK:0.26(0.60)对1.00(2.07),ULK1:0.27(3.31)对1.07(4.41),P值均<0.017],mTOR mRNA表达水平明显高于对照组[1.82(3.74)对1.26(1.38),P<0.017]。相对高危组MDS患者有核红细胞LC3B水平与HGB呈正相关(r=0.529,P=0.009)。相对高危组MDS患者有核红细胞TOM20蛋白(9.42±4.42)表达升高。 结论: 高危MDS患者有核红细胞自噬水平下降。.

Keywords: Anemia; Autophagy; Myelodysplastic syndromes.

MeSH terms

  • Autophagy*
  • Bone Marrow
  • Bone Marrow Cells
  • Erythroblasts
  • Erythrocyte Count
  • Erythrocytes
  • Flow Cytometry
  • Humans
  • Membrane Proteins
  • Microscopy, Electron, Transmission
  • Myelodysplastic Syndromes
  • TOR Serine-Threonine Kinases

Substances

  • Membrane Proteins
  • MTOR protein, human
  • TOR Serine-Threonine Kinases

Grants and funding

基金项目:国家自然科学基金(81170472、81400088);天津市应用基础与前沿技术研究计划(14JCYBJC27200、09JCYBJC11200);天津市抗癌重大专项攻关计划(12ZCDZSY17900)