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目的 :探讨索拉非尼对人多发性骨髓瘤细胞U266增殖和凋亡的影响,同时探索其作用机制。方法 :通过MTT法检测多发性骨髓瘤细胞U266在不同浓度的索拉非尼作用下的增殖抑制率;流式细胞术检测细胞凋亡率;同时设计引物VEGFR-2、VEGFR-3、PDGFR-β、c-Kit,采用Q-PCR检测索拉非尼处理后的多发性骨髓瘤细胞U266 m RNA表达水平,初步确定索拉非尼的作用靶点;最后用Western-blot检测PDGFR-β与VEGFR-3蛋白表达的情况。结果 :不同浓度的索拉非尼对多发性骨髓瘤细胞U266抑制增殖及促进凋亡作用各异,且抑制率和凋亡率均呈时间-浓度依赖性;索拉非尼作用48 h之后,与对照组比较VEGFR-2、VEGFR-3、PDGFR-β、c-Kit的m RNA表达均下调,其中VEGFR-3、PDGFR-β最为显著,Western-blot检测VEGFR-3、PDGFR-β蛋白表达量与Q-PCR结果一致。结论 :索拉非尼有抑制人多发性骨髓瘤细胞的增殖及促进其凋亡的作用,其机制可能与索拉非尼可以抑制细胞表面相关受体激活途径有关。
Objective: To investigate the effect of sorafenib on the proliferation and apoptosis of human multiple myeloma cell line U266, and to explore its mechanism. Methods: MTT assay was used to detect the proliferation inhibition rate of multiple myeloma cells U266 under different concentrations of sorafenib. Flow cytometry was used to detect the apoptosis rate. Meanwhile, the expression of VEGFR-2, VEGFR-3, PDGFR- β, c-Kit. The expression level of U266 m RNA in somatofenib-treated multiple myeloma cells was detected by Q-PCR and the target of sorafenib was determined. Western blotting was used to detect the expression of PDGFR- VEGFR-3 protein expression. RESULTS: Sorafenib at different concentrations had different effects on the proliferation and apoptosis of multiple myeloma cells U266 in a time-and concentration-dependent manner. After sorafenib treatment for 48 h, Compared with the control group, the mRNA expression of VEGFR-2, VEGFR-3, PDGFR-β and c-Kit were down-regulated, of which VEGFR-3 and PDGFR-β were the most significant Consistent with Q-PCR results. Conclusion: Sorafenib can inhibit the proliferation and promote the apoptosis of human multiple myeloma cells. The mechanism may be related to the inhibition of sorafenib on the activation of cell surface receptor.