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Ruxolitinib对人红白血病HEL细胞VEGF、HIF-1α表达的影响

The Effect of Ruxolitinib on the Expression of VEGF and HIF-1α in Leukemia HEL Cells

  • 摘要: 目的 探讨JAK2抑制剂Ruxolitinib对人红白血病HEL细胞血管内皮生长因子(VEGF)、缺氧诱导因子-1α(HIF-1α)分泌的影响。方法 用不同浓度Ruxolitinib(1、5、10、50、100、500 nmol/L)处理HEL细胞24、48、72 h,通过CCK-8法观察其对HEL细胞增殖的抑制作用;不同浓度Ruxolitinib处理细胞24、48、72 h,RT-PCR检测Jauns激酶2基因(JAK2) mRNA水平,Western blot检测p-JAK2、VEGF、HIF-1α蛋白表达;鸡胚绒毛尿囊膜(CAM)体内血管生长实验检测Ruxolitinib对血管生成的影响。结果 不同浓度Ruxolitinib(除外1 nmol/L作用24 h)均可抑制HEL细胞增殖。不同浓度Ruxolitinib处理HEL细胞24、48、72 h 后,RT-PCR结果显示JAK2 mRNA表达较对照组降低(P<0.01);Western blot结果显示p-JAK2、VEGF、HIF-1α蛋白表达较对照组亦均降低(P<0.05);CAM实验结果显示Ruxolitinib处理细胞72 h后血管数目明显减少。结论 Ruxolitinib可能通过抑制JAK2通路,抑制HEL细胞VEGF、HIF-1α表达进而抑制血管新生。

     

    Abstract: Objective To investigate the effect of Ruxolitinib on the expression of vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1 alpha (HIF-1α) in HEL cells. Methods The HEL cells were treated with Ruxolitinib in different concentrations (1 nmol/L, 5 nmol/L, 10 nmol/L, 50 nmol/L, 100 nmol/L, 500 nmol/L). The growth inhibition of Ruxolitinib on HEL cells was detected by CCK-8 assay; the mRNA expression level ofJAK2 were measured by RT-PCR and the protein level of p-JAK2, VEGF, HIF-1α were observed by Western blot after treated with Ruxolitinib for 24, 48, 72 h. Chick chorioallantoic membrane (CAM) test was used to testify the effect of Ruxolitinib on angiogenesis. Results Ruxolitinib with different concentrations could inhibit HEL cells proliferation. RT-PCR showed that the mRNA level ofJAK2 decreased in a concentration-dependent manner and Western blot demonstrated that the expression levels of p-JAK2, VEGF and HIF-1α were lower in Ruxolitinib treatment groups than those in control group (P <0.05) after HEL cells were treated with different concentrations of Ruxolitinib for 24, 48, 72 h. Ruxolitinib significantly suppressed blood vessels’ formation in CAM. Conclusion Ruxolitinib can inhibit VEGF, HIF-1α expression and angiogenesis of HEL leukemia cells by inhibiting JAK2 pathway.

     

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