摘要
目的探讨体外蛋白酶体抑制剂在肿瘤坏死因子相对凋亡诱导配体(TRAIL)诱导恶性淋巴瘤细胞凋亡抵抗中的作用及其分子机制。方法TRAIL与蛋白酶体抑制剂硼替唑咪(PS-341)处理恶性淋巴瘤Raji细胞后,采用MTF法检测药物不同作用时问对细胞的增生影响;流式细胞术检测细胞周期;Western blotting法检测Bax蛋白质水平变化;实时荧光定量RT—PCR法检测BaxmRNA的表达情况。结果TRAIL在质量浓度500μg/L时可减少Raji细胞的增生,但低于正常淋巴细胞Hmy2.ciR;TRAIL阻滞细胞周期于G0/G1期,Western blotting法检测Raji细胞Bax蛋白表达水平下降,实时荧光定量RT—PCR法检测BaxmRNA的表达水平却未见差异性改变,联合使用浓度为10nmol/LPS-341后可明显提高TRAIL的诱导凋亡作用,阻滞于G0/G1期的细胞数增加,Bax蛋白表达水平也逐渐增加。结论TRAIL在诱导恶性淋巴瘤细胞凋亡过程中出现抵抗,可能与促凋亡蛋白Bax表达水平下降有关,而Bax蛋白表达水平的下降与mRNA表达水平无关,可能与泛素-蛋白酶体途径的降解有关。
Objective To explore the effect and molecular mechanism of proteasome inhibitor in TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis resistance on malignant lymphoma cells. Methods Raji cells were treated with TRAIL and proteasome inhibitor (PS-341) in vitro and the cell growth index was evaluated by MTT assay; cell cycle was analysed by flow cytometry; the protein and mRNA level of Bax were measured by Western blotting and real time RT-PCR. Results TRAIL inhibited proliferation of Raji cells at the concentration of 500μg/L, but the inhibition rate was lower than that of the control cell: Hmy2.ciR.TRAIL arrested cell in G0/G1 phase. The Bax protein in Raji is degraded, but the Bax mRNA expression level does not change significantly.The effects of TRAIL was enhanced significantly 10 nmol/L PS- 341 was added. Conclusion Raji cells are resistant in TRAIL-induced apoptosis. This effect may be related to the decrease of Bax protein. The Ubiquitin-proteasome pathway is involved in the degradation of Bax in TRAIL-treated Raji cells.
出处
《白血病.淋巴瘤》
CAS
2009年第6期331-334,共4页
Journal of Leukemia & Lymphoma
基金
基金项目:国家自然科学基金(30600754)