摘要
目的探讨藤黄酸对多发性骨髓瘤细胞增殖及凋亡的影响,并探讨其分子学机制,为其临床治疗多发性骨髓瘤提供依据。方法用不同浓度的藤黄酸处理多发性骨髓瘤细胞株NCI-H929细胞,采用MTS法检测藤黄酸对多发性骨髓瘤细胞活性的影响;碘化丙啶单染法于荧光显微镜下观察细胞形态的变化;应用Western blot法检测泛素蛋白酶体系统相关蛋白(泛素-蛋白酶体系统蛋白(Ubs)、B细胞κ轻肽基因增强子核因子抑制因子α、B细胞淋巴瘤因子相关蛋白(Bax))、凋亡相关蛋白(聚腺苷二磷酸-核糖聚合酶(PARP)、含半胱氨酸的天冬氨酸蛋白水解酶3前体(Pro-caspase3)、髓细胞白血病-1蛋白(Mcl-1)、X连锁凋亡抑制蛋白(XIAP)、B细胞淋巴瘤-白血病-2蛋白(Bcl-2)及信号转导和转录激活因子5(STAT5))表达情况。结果藤黄酸明显抑制NCI-H929细胞的活力,诱导NCI-H929细胞凋亡;随藤黄酸浓度升高,Ubs聚集,Bax表达升高,蛋白酶体系统受到抑制;随藤黄酸浓度升高与作用时间延长,PARP出现切割,凋亡相关蛋白Pro-caspase3、Mcl-1、XIAP、Bcl-2及增殖通路蛋白STAT5表达下调。结论藤黄酸能够抑制多发性骨髓瘤细胞株NCI-H929的蛋白酶体活性,影响凋亡相关蛋白的表达,促进细胞凋亡。其机制可能通过抑制JAKs/STATs信号转导通路的活化,抑制多发性骨髓瘤细胞的增殖。
Objective To investigate the effects and its molecular mechanism of gambogic acid on cell proliferation and apoptosis multiple myeloma （MM） and provide basis for its therapy. Methods MM cell strain NCI-H929 cells were treated with different concentrations of gambogic acid. The impact of gambogie acid on MM eytoactive was detected by applying MTS assay. Cell morphological changes were observed by inverted microscope and fluorescence microscope with PI staining. The changes of protea some proteins including ubiquitin-proteasome system protein （Ubs）, nuclear factor κB inhibitor protein （κB-α）, Bcl-2 associated X protein （Bax）, apoptosis proteins including Poly ADP-ribose polymerase （PARP）, precursors of cysteinyl aspartate specific pro- teinase 3 （Pro easpase3）, myeloid cell leukemia 1 （Mcl-1）, X-linked inhibitor of apoptosis protein （XIAP）, B cell lymphoma/leu- kemia-2 （Bcl-2） and proliferation associated proteins including signal transduction and activator of transcription 5 （STATS） were tested by Western blot method. Results Garnbogie acid markedly inhibited the cytoaetive of NCI-H929 cells and induced apopto- sis. Along with the increase of gambogic acid concentration, Ubs protein accumulated, Bax expression elevated, and proteases were inhibited. Along with the increase of the gambogic acid concentration and extension of its action time, cleavage of PARP appeared, apoptosis-associated proteins Pro easpase3, Mcl-1, XIAP, Bcl 2 and the expression of proliferation pathway-related protein STAT5 downregulated. Conclusion Gambogic acid can inhibit the proteasome activity in multiple myeloma cell line NCI H929, impact the expression of apoptosis related proteins and advance apoptosis. The mechanism is likely through restraining the activation of JAKs/STATs signal transduction pathway and thus inhibiting cell proliferation of this disease.
出处
《青岛大学医学院学报》
CAS
2013年第5期408-411,共4页
Acta Academiae Medicinae Qingdao Universitatis
