基础医学与临床 ›› 2014, Vol. 34 ›› Issue (2): 190-195.

• 研究论文 • 上一篇    下一篇

PTEN对髓系白血病细胞VEGF及MMP表达的影响

成志勇1,王亚丽2,王凤云2,韩英2,谷蕾2,梁丽青2,谢旭磊2   

  1. 1. 保定市第一医院血液内科
    2. 保定市第一医院
  • 收稿日期:2013-06-09 修回日期:2013-10-17 出版日期:2014-02-05 发布日期:2014-01-13
  • 通讯作者: 成志勇 E-mail:dzczy@sohu.com
  • 基金资助:
    河北省自然科学基金资助项目:PTEN基因在慢性粒细胞性白血病中的作用及其机制的研究。

The influence of of PTEN on VEGF and MMP expression in myeloid leukemia cells

  • Received:2013-06-09 Revised:2013-10-17 Online:2014-02-05 Published:2014-01-13
  • Contact: CHENG Zhi Yong E-mail:dzczy@sohu.com

摘要: 目的 探讨在慢性髓系白血病中与张力蛋白同源的10号染色体缺失的磷酸酶基因(PTEN)对血管内皮生长因子(VEGF)及金属基质蛋白酶(MMP)相互影响及其作用机制。方法 1)研究10例慢性髓系白血病慢性期CML-CP、10例急变期CML-BC及10例正常人骨髓单个核细胞内PTEN、VEGF、MMP-2和MMP-9 mRNA表达水平变化。2)将携带有野生型PTEN和绿色荧光蛋白的腺病毒(Ad-PTEN-GFP)及对照载体腺病毒(Ad-GFP)转染人慢性髓系白血病细胞系K562。MTT检测细胞增殖;荧光定量PCR(FQ-PCR)检测PTEN、VEGF、MMP-2和MMP-9 mRNA水平变化,Western blot及明胶酶谱检测PTEN、p-Akt、VEGF、MMP-2和MMP-9蛋白表达。结果 CML-BC患者中PTEN mRNA表达水平低于CML-CP及正常对照组(P<0.05),而VEGF、MMP-2、MMP-9 mRNA在CML-BC患者中均高于CML-CP及正常对照组(P<0.05)。以MOI=200转染K562细胞3d后Ad-PTEN-GFP组K562细胞内VEGF、MMP-2、MMP-9 mRNA表达水平均明显低于Ad-GPF组和未转染组(P<0.05),PTEN与VEGF、MMP-2、MMP-9 mRNA表达水平负相关,转染Ad-PTEN-GFP组与转染Ad-GFP组比较VEGF、MMP-2、MMP-9 mRNA表达水平分别降低4.80、5.88、5.72倍,p-Akt及VEGF、MMP-2、MMP-9蛋白表达水平分别降低26.0、5.23、2.86、4.76倍。结论 PTEN基因可能通过抑制VEGF、MMP-2、MMP-9表达,抑制髓系白血病细胞血管新生及侵袭。

Abstract: Objective To investigate the influence and relationship of tumor-suppressing gene PTEN (phosphatase and tensin hemology deleted on chromosome ten gene), VEGF(vascular endothelial growth factor) and MMP (matrix metalloproteinase) in myeloid leukemia. Methods 1)To explore the PTEN, VEGF, MMP-2 and MMP-9 mRNA expression levels on 10 chronic myelogenous leukemia patients in chronic phase (CML-CP), 10 CML patients in blast crises (CML-BC), 10 normal control marrow mononuclear cells (MMNC). 2)The recombinated adenovirus containing green fluorescent protein (GFP) and PTEN(Ad-PTEN-GFP)or empty vector (Ad-GFP)was transfected into human leukemia K562 cells. The growth inhibition ratio of K562 cells were observed by MTT assay; PTEN, VEGF, MMP-2 and MMP9 mRNA levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR (FQ-PCR). PTEN, VEGF and p-Akt protein levels were detected by western blot, MMP-2 and MMP-9 protein were detected by gelatin zymogram. Results: The mRNA expression levels of PTEN in CML-BC patients were lower than CML-CP patients(P<0.05). But VEGF, MMP-2 and MMP-9 mRNA levels were reverse compared with PTEN(P<0.05). VEGF, MMP-2 and MMP-9 mRNA expression levels were down regulated about 4.80、5.88、5.72 fold; p-Akt, VEGF, MMP-2 and MMP-9 proteins were also down-regulated 26.0, 5.23, 2.86, 4.76 fold compared with Ad-GFP group; PTEN mRNA expression levels were negative with VEGF, MMP-2, MMP-9 mRNA after K562 cells transfected with wild type PTEN of MOI=200 after three days. Conclusion PTEN might have anti-angiogenesis and anti-adhesion ability in myeloid leukemia via inhibition VEGF, MMP-2 and MMP-9 expression.