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表达水平呈负相关,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79.12%,82.99%,82.52%,p-Akt及VEGF、MMP-2、MMP-9蛋白表达水平分别降低96.15%,80.88%,65.03%,78.99%。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。
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 （ma- trix metalloproteinase） in myeloid leukemia. Methods 1 ） To explore the PTEN, VEGF, MMP-2 and MMP-9 mR- NA expression 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 contai- ning 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 MTI＂ assay; PTEN, VEGF, MMP-2 and MMP-9 mRNA levels were detected by real-time fluorescent relative-quantification reverse tran- scriptional PCR （FQ-PCR）. PTEN, VEGF and p-Akt protein levels were detected by western blot,MMP-2 andMMP-9 protein were detected by gelatin zymogram. Results The mRNA expression of PTEN in CML-BC patients was lower than that in CML-CP patients（P 〈 0. 05 ）. But VEGF, MMP-2 and MMP-9 mRNA levels were reverse compared with PTEN （ P 〈 O. 05 ）. VEGF, MMP-2 and MMP-9 mRNA expression levels were down regulated about 79. 15% ,82. 99%, 82. 52% ; p-Akt, VEGF, MMP-2 and MMP-9 proteins were also down-regulated 96. 15%, 80. 88% ,65.03% ,78.99% compared with Ad-GFP group; PTEN mRNA expression was negative with VEGF, MMP-2, MMP-9 mRNA after K562 cells transfected with wild type PTEN of MOI = 200 after three days. Conclu- sions PTEN might have anti-angiogenesis and anti-adhesion ability in myeloid leukemia via inhibition VEGF, MMP-2 and MMP-9 expression.
出处
《基础医学与临床》
CSCD
北大核心
2014年第2期190-195,共6页
Basic and Clinical Medicine
基金
河北省自然科学基金(C2010000538)
