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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、500nmol/L)处理HEL细胞24、48、72h,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、72h,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1nmol/L作用24h)均可抑制HEL细胞增殖。不同浓度Ruxolitinib处理HEL细胞24、48、72h后,RT-PCR结果显示JAK2mRNA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72h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。
Objective To investigate the effect of JAK2 inhibitor Ruxolitinib on the secretion of vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1α (HIF-1α) in human erythroleukemia cell line HEL. Methods HEL cells were treated with different concentrations of Ruxolitinib (1, 5, 10, 50, 100 and 500 nmol / L) for 24 h, 48 h and 72 h, respectively. The inhibitory effects of Ruxolitinib on HEL cell proliferation were observed by CCK- , 48 and 72h respectively. The mRNA expression of JAK2 was detected by RT-PCR and the protein expression of p-JAK2, VEGF and HIF-1α were detected by Western blot. The in vivo growth of chick chorioallantoic membrane was detected by Ruxolitinib Effects of angiogenesis. Results Ruxolitinib with different concentrations (all except 1nmol / L for 24h) could inhibit the proliferation of HEL cells. After treated with Ruxolitinib for 24,48,72 h, the expression of JAK2 mRNA in HEL cells was lower than that in control group (P <0.01). Western blot results showed that the expression of p-JAK2, VEGF and HIF-1α (P <0.05). The results of CAM assay showed that the number of blood vessels in Ruxolitinib treated cells decreased significantly after 72 hours. Conclusion Ruxolitinib may inhibit angiogenesis by inhibiting the JAK2 pathway and inhibiting the expression of VEGF and HIF-1α in HEL cells.