三种人多发性骨髓瘤细胞系移植小鼠模型的建立和比较

Establishment and comparison of three human multiple myeloma cell line transplantation models in mice

目的利用人骨髓瘤细胞系ARP1、MM.1S和NCI-H929建立异种移植模型, 并对三种细胞移植后生长周期、肿瘤负荷和生物学特点进行比较。方法将ARP1、MM.1S和NCI-H929细胞分别由皮下或尾静脉植入经137Cs照射后的NOD/SCID小鼠, 每周观察小鼠的生存情况, 监测肿瘤负荷。应用流式细胞术检测小鼠肿瘤组织或骨髓中CD138+细胞的比例。采用免疫荧光检测肿瘤组织细胞的CD138和免疫球蛋白轻链表达。ELISA法检测骨髓和外周血中的免疫球蛋白轻链, micro-CT评价骨病变。结果皮下移植ARP1、MM.1S和NCI-H929细胞的小鼠在两周内均可形成局部肿瘤, 免疫荧光检测支持浆细胞肿瘤。尾静脉移植后第20天时可在ARP1组小鼠外周血中检测出κ轻链[(8.2±1.0)ng/ml]。尾静脉移植6周左右, ARP1组小鼠出现体重下降、精神萎靡、下肢逐渐瘫痪等表现, 骨髓中能检测到人CD138+CD38+细胞群, 予硼替佐米治疗可显著降低肿瘤负荷[(5.7±0.2)%对(21.3±2.1)%, P<0.01]。MM.1S和NCI-H929组小鼠骨髓中未检测到人CD138+CD38+细胞群。结论 ARP1、MM.1S和NCI-H929细胞系构建的小鼠模型可作为MM发病机制及临床研究的良好模型。

Objective To establish three types of xenotransplantation models using human myeloma cell lines ARP1,MM.1S,and NCI-H929 and to compare the proliferation,tumor load,and biological characteristics of the three types of cells after transplantation.Methods Suspensions of human myeloma cell lines ARP1,MM.1S,and NCI-H929 were implanted into NOD/SCID mice by subcutaneous injection or tail vein injection.The survival of the mice was observed weekly,and the tumor load was measured.Flow cytometry was used to detect the proportion of CD138+cells in tumor tissue or the mouse bone marrow.CD138+cells and light chains were detected by immunofluorescence.Light chains in bone marow and peipheral blood were measured by ELISA,and bone disease was assessed by micro-CT.Results Mice injected with ARP1,MM.1S,and NCI-H929 cells all formed tumors subcutaneously in about 2 weeks.Immunofluorescence detection supported plasma cell tumors.Kappa light chains were detected in the peripheral blood of ARP1 mice on day 20 after tail vein transplantation(8.2±1.0 ng/ml).After 6 weeks of tail vein transplantation,mice in the ARP1 group showed signs of weight loss,mental depression,and dragging legs,and human CD138+CD38+cells were detected in the bone marrow(BM).Furthermore,bortezomib(BTZ)treatment given once the tumor was established significantly reduced the tumor burden[(5.7±0.2)%vs(21.3±2.1)%,P<0.01].Human CD138+CD38+cells were not detected in the BM of the MM.1S or NCI-H929 groups.Conclusion The results of this study suggest that the mouse models constructed by the three cell lines(ARP1,MM.1S,and NCI-H929)can be used as models for the pathogenesis and clinical research of MM.