多西环素对骨髓瘤细胞株H929内源性凋亡的影响及其作用机制  

Effect of Doxycycline on Intrinsic Apoptosis of Myeloma Cell Line H929 and Its Mechanism

作　　者：李海璐 费小明[1] 汤郁[2] 杨元林 王丽霞[1] 耿佳伟 LI Hai-Lu;FEI Xiao-Ming;TANG YU;YANG Yuan-Lin;WANG Li-Xia;GENG Jia-Wei(Depariment of Hematology,Zhenjiang 212000,Jiangsu Province,China;Deparment of Rheumatology,The Affliated Hospital of Jiangsu University,Zhenjiang 212000,Jiangsu Province,China)

机构地区：[1]江苏大学附属医院血液科,江苏镇江212000 [2]江苏大学附属医院风湿科,江苏镇江212000

出　　处：《中国实验血液学杂志》2022年第2期441-448,共8页Journal of Experimental Hematology

基　　金：江苏省卫生健康委科研课题(H2018084);江苏省社会发展重点项目(临床前沿技术)课题(BE2020681);国家自然科学基金(81571582)。

摘　　要：目的:探索多西环素(DOX)诱导骨髓瘤H929细胞株凋亡的作用及其可能机制。方法:用DOX、MEK抑制剂U0126和RAS激动剂ML-098单独或联合处理H929细胞,Western blot检测p-MEK、caspase-3、caspase-9、c-Jun等蛋白的表达;CCK8法检测细胞增殖;Annexin-V-FITC/PI双染法检测细胞凋亡。结果:经DOX处理H929细胞后,p-MEK蛋白表达明显减少(P<0.05),伴有caspase-3、9蛋白剪切体的明显增加(P<0.05)。U0126处理H929细胞后,p-MEK蛋白水平明显下降(P<0.05),伴有caspase-3、9蛋白剪切体的明显增加(P<0.05)。DOX联合ML-098处理H929细胞后,H929细胞凋亡比例较DOX单独处理组减少;两药联合处理组p-MEK及p-Erk1/2蛋白表达明显高于DOX单独处理组(P<0.05), Caspase-3、9蛋白剪切体的水平较DOX单药处理组均明显下降(P<0.05)。DOX处理H929细胞后,c-Jun mRNA和蛋白表达水平均明显增加(P<0.05)。结论:DOX可诱导H929细胞内源性凋亡途径的激活,而MEK/ERK通路及c-Jun可能参与DOX诱导细胞凋亡。Objective: To investigate the mechanism of the in vitro toxicity of doxycycline to myeloma cell line H929 and also the possible pathway involved its toxicity. Methods: Myeloma cell line H929 was treated with DOX, MEK inhibitor U0126 or RAS agonist ML-098, either alone or in combination. Then, the expression of p-MEK, caspase-3, caspase-9 and c-Jun in H929 were used to detected by Western blot;the cells proliferation and apoptosis were detected by CCK-8 assay and flow cytometry, respectively. Results: DOX significantly increased the levels of cleaved caspase-3 and caspase-9, and down-regulated the level of p-MEK in H929(P<0.05). MEK antagonist U0126 significantly increased the levels of cleaved caspase-3 and caspase-9, and down-regulated the level of p-MEK(P<0.05). After Dox combined with ML-098 treatment of H929 cells, the apoptosis rate of H929 cells was lower than that of DOX alone treatment group(P<0.05). Compared with DOX alone treatment group, the expressions of p-MEK and p-ERK1/2 in DOX+ML-098 combined treatment group were increased, and the levels of cleaved caspase-3,9 in H929 cells were decreased(P<0.05). The levels of c-Jun mRNA and protein increased in H929 when treated by DOX alone(P<0.05). Conclusion: DOX can induce apoptosis of H929 via intrinsic apoptosis pathway, and MEK/ERK pathway and c-Jun possibly play a role in this process.

关 键 词：多发性骨髓瘤 多西环素 H929细胞 凋亡 MEK/ERK C-JUN 

分 类 号：R733.3[医药卫生—肿瘤]