利妥昔单抗与依鲁替尼对弥漫性大B细胞淋巴瘤细胞增殖和凋亡协同作用研究

发布时间：2018-06-14 00:18

本文选题：利妥昔单抗 + 依鲁替尼　；参考：《中华肿瘤防治杂志》2017年08期

【摘要】：目的近年来发现依鲁替尼是B细胞恶性肿瘤的新型靶向药物,利妥昔单抗联合新药依鲁替尼对弥漫性大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)的研究逐步展开,本研究探讨利妥昔单抗与依鲁替尼对DLBCL细胞系Farage增殖及凋亡影响的协同作用。方法不同浓度的单药利妥昔单抗和单药依鲁替尼处理Farage细胞24、48和72h后,采用CCK8法检测细胞增殖抑制率。15μmol/L依鲁替尼处理Farage细胞48h后,采用RT-PCR法检测凋亡相关因子mRNA的表达变化。含人补体的培养条件下,1μg/mL利妥昔单抗联合10μmol/L依鲁替尼同时处理Farage细胞48h后,采用CCK8法检测增殖抑制率,采用流式细胞术检测凋亡率。结果利妥昔单抗在不含人补体的培养基中对Farage细胞增殖抑制作用不明显,在含人补体的培养基中有明显增殖抑制作用,并呈浓度和时间依赖性。依鲁替尼在含或不含人补体的培养条件下对Farage细胞均有明显增殖抑制作用,呈浓度和时间依赖性,2种条件下的增殖抑制率差异无统计学意义(F=0.978,P=0.329),15μmol/L依鲁替尼作用Farage细胞48h时,Fas(1.63±0.09)、Caspase-8(1.90±0.11)和Caspase-3(2.20±0.11)mRNA相对表达量均高于空白组(1.00±0.00)。在含人补体的培养基中,1μg/mL利妥昔单抗联合10μmol/L依鲁替尼同时处理Farage细胞48h后,联合组增殖抑制率为(57.06±1.48)%,高于依鲁替尼单药组(33.83±3.39)%和利妥昔单抗单药组(26.92±2.74)%,差异有统计学意义,F=87.403,P0.001。联合组的凋亡和坏死率(44.30±2.20)%高于利妥昔单抗单药组(21.73±2.00)%和依鲁替尼单药组(17.44±1.60)%,且利妥昔单抗单药组和依鲁替尼单药组均高于空白组(2.32±0.21)%,差异均有统计学意义,F=327.205,P0.001。结论利妥昔单抗可通过CDC效应引起DLBCL细胞凋亡坏死及增殖抑制,依鲁替尼可能通过上调Fas、Caspase-3和Caspase-8基因的表达,促使DLBCL细胞凋亡和增殖抑制,利妥昔单抗联合依鲁替尼对DLBCL细胞的增殖抑制和凋亡坏死作用明显强于单药利妥昔单抗或依鲁替尼。
[Abstract]:Objective in recent years, it has been found that ilutinib is a new target drug for B cell malignant tumor. The study of rituximab combined with new drug ilutinib on diffuse large B cell lymphomavirus (DLBCLs) has been carried out step by step. The aim of this study was to investigate the synergistic effects of rituximab and ilutinib on the proliferation and apoptosis of DLBCL cell line Farage. Methods Farage cells were treated with different concentrations of Rituximab and irutinib for 24 h and 72 h, respectively. The inhibitory rate of proliferation of Farage cells was measured by CCK8 assay. The expression of apoptosis-related factors mRNA was detected by RT-PCR after treatment of Farage cells with 15 渭 mol / L ilutinib for 48 h. Farage cells were treated with 1 渭 g / mL Rituximab and 10 渭 mol / L ilutinib for 48 h under the condition of human complement. The inhibition rate of proliferation was measured by CCK8 method, and the apoptosis rate was detected by flow cytometry. Results the inhibitory effect of Rituximab on the proliferation of Farage cells was not obvious in the medium without human complement, but in the medium containing human complement in a concentration-and time-dependent manner. Yirutinib could inhibit the proliferation of Farage cells in culture with or without human complement. There was no significant difference in the inhibition rate of proliferation between the two groups in a concentration and time dependent manner. The relative expression levels of FasN and Caspase-32.20 卤0.11 mRNA in Farage cells were 1.63 卤0.09 Caspase-8N 1.90 卤0.11) and 1.00 卤0.000.11 at 48h after treatment with Yirutinib at 15 渭 mol 路L ~ (15) 渭 mol 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1). After 48 hours of treatment of Farage cells with 1 渭 g / mL Rituximab and 10 渭 mol / L ilutinib simultaneously, the inhibitory rate of proliferation in the combined group was 57.06 卤1.48, which was higher than that in the rituximab group (33.83 卤3.39U%) and the Rituximab group (26.92 卤2.74g / L). The difference was statistically significant (P 0.001). The apoptotic and necrotic rate of the combined group was 44.30 卤2.20% higher than that of the Rituximab group (21.73 卤2.00) and the rutinib group (17.44 卤1.60), and the Rituximab group and the ilutinib group were higher than that of the control group (2.32 卤0.21). The difference was statistically significant. Conclusion Rituximab can induce apoptosis, necrosis and proliferation inhibition of DLBCL cells through CDC effect. Elutinib may promote apoptosis and proliferation inhibition of DLBCL cells by up-regulating the expression of Fas-caspase-3 and Caspase-8 genes. The inhibitory effect of rituximab combined with irutinib on the proliferation and apoptosis of DLBCL cells was significantly stronger than that of Rituximab or elutinib.
【作者单位】：广西医科大学附属肿瘤医院淋巴血液肿瘤科;
【基金】：国家自然科学基金(81160298) 广西医疗卫生适宜技术研究与开发课题(S201301-02)
【分类号】：R733.1

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