硼替佐米联合多柔比星对淋巴母细胞淋巴瘤Molt-4细胞的作用

 目的　研究硼替佐米联合多柔比星对淋巴母细胞淋巴瘤细胞株Molt-4的作用。方法　硼替佐米与多柔比星作用于Molt-4细胞后，采用CCK-8检测细胞的增殖，锥虫蓝染色计数细胞的活性，应用AnnexinV-FITC凋亡试剂盒及线粒体膜电位检测试剂盒经流式细胞术（FCM）检测细胞凋亡，用免疫标记经FCM检测细胞膜表面Fas的表达。结果　硼替佐米与多柔比星单药作用对Molt-4细胞具有生长抑制作用，生长抑制率与药物浓度呈正相关（r＝0.863，P＝0.027；r＝0.915，P＝0.038）；两者联合应用对细胞生长抑制有协同作用，48 h作用最明显，抑制率达（57.24±0.10）%；联合应用细胞总凋亡率增高48 h达（23.08±1.25）%，与硼替佐米和多柔比星单药组（15.11±1.09）%和（15.96±0.81）%比较差异具有统计学意义（t值分别为0.046，0.037，均P＜0.05），早期凋亡率无明显变化（t值分别为0.052、0.048，均P＞0.05）。线粒体膜电位检测显示联合作用较单药作用细胞凋亡率升高分别为15.84 %、5.38 %、5.52 %，FCM检测药物联合作用后细胞表面Fas表达无明显改变（t＝0.040，P＞0.05）。结论　硼替佐米联合多柔比星能够有效抑制淋巴母细胞淋巴瘤细胞株Molt-4细胞增殖并诱导其凋亡，作用机制与活化线粒体凋亡途径有关，与外源性凋亡途径无关。

Effect of bortezomib combined with doxorubicin on lymphoblastic lymphoma Molt-4 cells

Objective　To investigate the effect of bortezomib combined with doxorubicin on lymphoblastic lymphoma cell line Molt-4. Methods　Molt-4 cells were cultured in the presence of bortezomib and doxorubicin, cell viability was monitored by CCK-8 and trypan-blue exclusion. Apoptosis was detected by flow cytometry and mitochondrial membrane potential, expression of Fas was also measured with flow cytometry. Results　Molt-4 cell proliferation was substantially inhibited in concentration-dependent manners when treated with either bortezomib or doxorubicin. The combination of both drugs synergistically inhibited Molt-4 cell proliferation at 48 hours [(57.24±0.10) %]. Combination therapy further enhanced bortezomib and doxorubicin induced apoptosis [48 h (23.08±1.25) %] (P ＜ 0.05). Detection of mitochondrial membrane potential showed that combination therapy could promote apoptosis (15.84 %, 5.38 %, 5.52 %) but did not significantly change the level of Fas expression (P ＞ 0.05). Conclusion　Combination therapy of bortezomib with doxorubicin efficiently inhibits proliferation and induces apoptosis of Molt-4 cells. Activation of mitochondrial and intrinsic apoptotic pathway may play important roles.