补肾养骨汤对人多发性骨髓瘤耐药细胞KM3/BTZ多药耐药性的影响  

Effect of Bushen Yanggu Decoction on Multidrug Resistance of Multiple Myeloma Cell Line KM3/BTZ

作　　者：李玉鹏 张建国 方梓青 黄幸儒 陈旭 李振杰 方坚 LI Yu-Peng;ZHANG Jian-Guo;FANG Zi-Qing;HUANG Xing-Ru;CHEN Xu;LI Zhen-jie;FANG Jian(The Third Clinical Medical School of Guangzhou University of Tranditional Chinese Medicine;Department of Orthopedics The Third Affiliated Hospital of Guangzhou University of Tranditional Chinese Medicine;Guangzhou Liwan District Orthopedics Hospital,Guangzhon 510000,Guangdong Province,China)

机构地区：[1]广州中医药大学第三临床医学院 [2]广州中医药大学第三附属医院骨科 [3]广州市荔湾区骨伤科医院,广东广州510000

出　　处：《中国实验血液学杂志》2019年第2期482-488,共7页Journal of Experimental Hematology

基　　金：广东省科技计划项目(20164009);广东省中医药局(重点项目)(20164009)

摘　　要：目的:探讨补肾养骨汤对人多发性骨髓瘤耐药细胞KM3/BTZ耐药性及增殖的影响,为补肾养骨汤在骨髓瘤治疗方面的前景和应用提供参考。方法:采用硼替佐米(bortezomib,BTZ)浓度梯度诱导法建立人多发性骨髓瘤耐药细胞KM3/BTZ,通过MTT法检测常用化疗药物以及含补肾养骨汤药血清对KM3细胞和KM3/BTZ细胞增殖的影响,采用RT-qPCR及Western blot法检测Par-4、HSP27和P-gp基因表达变化,采用流式细胞术检测细胞凋亡情况。结果:建立的KM3/BTZ细胞对常用化疗药物均可产生不同程度的耐药性,其中BTZ的耐药指数(RI)最高,为20.269。MTT法检测结果显示,经含补肾养骨汤药血清处理过的KM3/BTZ细胞增殖受抑制,且随含补肾养骨汤药血清浓度升高抑制作用呈增强趋势。含补肾养骨汤药血清可抑制KM3/BTZ细胞增殖,诱导其凋亡,明显降低KM3/BTZ细胞耐药性,上调Par-4表达,下调HSP27和P-gp表达。结论:补肾养骨汤能有效抑制KM3/BTZ细胞增殖,诱导其凋亡,逆转KM3/BTZ细胞多药耐药性,其机制可能与HSP27和P-gp表达下调,Par-4表达升高有关。Objective:To explore the effect of Bushen Yanggu Decoction(BYD)on drug resistance and proliferation of human multiple myeloma-resistant KM3/BTZ cells.Methods:Human multidrug-resistant KM3/BTZ cells were established by Bortezomib(BTZ)gradient induction.The effects of commonly used chemotherapeutic drugs and serum containing Bushen Yanggu Decoction(BYD)on the proliferation of KM3 cells and KM3/BTZ cells were detected by MTT assay.RT-qPCR and Western blot were used to detect the expression of Par-4,HSP27 and P-gp genes.Flow cytometry was used to detect cell apoptosis.Results:The established KM3/BTZ cells could produce varying degree of resistance to commonly used chemotherapeutic drugs.Among them,the highest resistance index(RI)to BTZ was 20.269.MTT assay showed that the proliferation of KM3/BTZ cells treated with serum containing Bushen Yanggu Decoction was inhibited,and the inhibitory effect increased with the serum concentration incranse of Bushen Yanggu Decoction.The serum containing Bushen Yanggu Decoction could inhibit the proliferation of KM3/BTZ cells,and induce apoptosis,significantly reduce the drug-resistance of KM3/BTZ cells,up-regulate the expression of Par-4,down-regulate the expression of HSP27 and P-gp.Conclusion:Bushen Yanggu Decoction can effectively inhibit the proliferation of KM3/BTZ cells and induce apoptosis.Bushen Yanggu Decoction can effectively reverse the multidrug-resistance of KM3/BTZ cells.The mechanism may be related with the decrease of expression of HSP27 and P-gp and the increase of expression of Par-4.

关 键 词：补肾养骨汤 多发性骨髓瘤 硼替佐米 多药耐药 

分 类 号：R733.3[医药卫生—肿瘤]