白血病患者骨髓中CTGF、CYR61、VEGF-C、VEGFR-2mRNA、蛋白的表达及其临床相关性研究

本研究旨在探讨白血病患者骨髓单个核细胞中结缔组织生长因子（CTGF）、富含半胱氨酸蛋白61（CYR61）、血管内皮生长因子-c（VEGF—C）和血管内皮生长因子受体-2（VEGFR-2）mRNA及蛋白的表达水平以及它们在白血病发生、发展、浸润转移中的作用和临床意义。初发白血病100例患者，26例完全缓解后急性白血病患者，30例对照者纳入本研究。收集骨髓单个核细胞，应用realtimePCR和Westernblot分别检测白血病患者及对照者CTGF、CYR61、VEGF-C、VEGFR-2mRNA及蛋白的表达水平。结果表明，急性白血病患者骨髓中上述4种因子mRNA、蛋白水平明显高于对照组（P〈0．05）。急性白血病缓解后只有CTGFmRNA表达与对照组比较有差异（P〈0．05），且CTGFmRNA表达在急性白血病不同骨髓增生中有统计学意义（P〈0．05）。CTGF蛋白表达在白血病不同染色体核型组有差异（P〈0．05），CYR61和VEGF—C蛋白表达在急性白血病不同程度骨髓增生组中比较差异有统计学意义（P〈0．05）。CML组CTGF、CYR61、VEGF—CmRNA和蛋白表达高于对照组（P〈0．05），CLL组CTGF和CYR61蛋白表达水平高于对照组。在性别、髓外浸润等因素分组比较中，上述4种因子mRNA和蛋白表达水平比较均无差异有统计学意义（P〉0．05）。在相关性分析中，白血病患者上述4种因子mRNA表达水平与骨髓原始细胞计数呈正相关（P〈0．05），CTGF、CYR61、VEGF—C蛋白表达水平与其呈正相关（P〈0．05）。除了VEGFR-2与骨髓原始细胞计数无相关性外，这些因子的mRNA及蛋白表达水平与骨髓原始细胞计数呈正相关。结论：CYR61、CTGF、VEGF-C和VEGFR-2mRNA和蛋白表达在急慢性白血病中都有作用。在急性白血病（AML／ALL）中上述4种因子高表达，在CML中除了VEGFR-2外其他因子也高表达，多数因子的表达与骨髓原始细胞计数呈正相关。联合阻断这些相关的血管性因子可能对白血病靶向治疗有重要作用。

Protein and mRNA Expression of CTGF,CYR61, VEGF-C and VEGFR-2 in Bone Marrow of Leukemia Patients and Its Correla- tion with Clinical Features

This study was aimed to investigate the mRNA and protein expression of CTGF, CYR61, VEGF-C and VEGFR-2 in bone marrow of patients with leukemia, and to analyze the role and clinical significance of these 4 factors in genesis and development of leukemia, infiltration and metastasis of leukemic cells. A total of 100 cases of newly diagnosed leukemia, 26 cases of acute leukemia in complete remission and 30 controls were enrolled in this study. The mononuclear cells of bone marrow were collected, the mRNA and protein expression levels of CTGF, CYR61, VEGF- C, VEGFR-2 in leukemia patients and controls were detected by real time PCR and Western blot, resoectivelv. The re-suits showed that the mRNA and protein expression levels of above mentioned 4 factors were significantly higher than those in control （ P 〈 0.05 ）, only CTGF mRNA expression in AL patients after complete remission showed statistical difference as compared with control（P 〈 0.05 ）, but the expression of CTGF mRNA showed statistical significance in different bone marrow hyperplasia of acute leukemia （ P 〈 0.05 ）. The expression level of CTGF protein showed difference in different chromosome karyotypes of leukemia （ P 〈 0.05 ）. The expression levels of CYR61 and VEGF-C proteins showed statistical difference in different bone marrow hyperplasia of acute leukemia （ P 〈 0.05 ）. The expression level of CTGF,CYR61, VEGF-C mRNA and protein in CML group were higher than that in control group. The expression levels of CTGF and CYR61 patein were higher than that in control. The mRNA and protein expression levels of above-menthioned 4 factors in sex and infiltration If leukemic cells did not show statistical significance （ P 〈 0.05 ）. In correlative analysis, the mRNA expressions of above menthoned 4 factors were possitively correlated with bone marrow blas count（ P 〈 0.05 ）, the protein expression of CTGF, CYR61 and VEGF-C were possitively correlated with bone marrow blast count. It is concluded that the CTGF, CYR61, VEGF-C and VEGFR-2 mRNA and protein play a role in acute leukemia. In acute leukemia （AML/ALL）, the expression of above menthined factor was high, but exept VEGFR-2. Most of them were positively correlated with bone marrow blast count. Joint block of these angiogenesis-related factors is likely to play an important role in targeting treatment of leukemia.