[Analysis of Tumorigenesis in Mice Transplanted with Different Multiple Myeloma Cell Lines]

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2019 Oct;27(5):1522-1529. doi: 10.19746/j.cnki.issn.1009-2137.2019.05.025.
[Article in Chinese]

Abstract

Objective: To investigate the tumorigenicity of several multiple myeloma (MM) cell lines transplanted in mice without γ-ray irradiation and to construct the MM disease model to facilitate in vivo experiments.

Methods: NOD/SCID or NSG mice were subcutaneously or caudally transplanted with MM cell lines (LP-1, OPM2, RPMI 8226 and MOLP8), or cell lines with luciferase (RPMI-Luc-Puro, RPMI-Luc-mCherry and MOLP8-Luc-Puro). Tumor growth was observed by measuring the tumor size with a caliper. CD138+ tumor cells in peripheral blood were detected by flow cytometry. The free light chain in mouse serum was detected by immunofixation electrophoresis. Tumor type was identified by immunohistochemistry.

Results: Twenty one NOD/SCID mice were subcutaneously transplanted with LP-1 cells or OPM2 cells respectively, and no tumors formed till 7 weeks after transplantation. Fifteen NOD/SCID mice subcutaneously transplanted with RPMI 8226 cells showed tumor formation one week later. As of 7 weeks, the rate of tumorigenesis was 80% (12/15). Serum λ light chain was detected and no CD138+ tumor cells were detected in peripheral blood. Two NOD/SCID mice each were subcutaneously transplanted with RPMI-Luc-Puro, RPMI-Luc-mcherry and MOLP8-Luc-Puro cells respectively. No tumor signal was detected through IVIS in RPMI-Luc-mcherry cells-transplanted mice. There was tumor signal at 1 week in RPMI-Luc-Puro and MOLP8-Luc-Puro cells-transplanted mice, the former disappeared at 2 weeks and the latter persisted more than 3 weeks. NSG mice subcutaneously transplanted with both cells persistently displayed the tumor signal. Neither NOD/SCID nor NSG mice transplanted with RPMI 8226, RPMI-Luc-Puro, RPMI-Luc-mcherry or MOLP8-Luc-Puro cells through tail vein developed the tumor signal. Only one NSG mice transplanted with MOLP8-Luc-Puro cells appeared transient tumor signal.

Conclusion: Unirradiated mice transplanted with MM cell lines tended to develop local tumor, and failed to develop disseminated tumor. The tumorigenicity of different cell lines is quite different and the vector transfection can reduce the tumorigenic ability. NSG mice with more severe immunodeficiency are more suitable for tumor growth.

题目: 不同多发性骨髓瘤细胞系移植小鼠的成瘤情况分析.

目的: 探讨在无γ射线照射条件下,几种常见的多发性骨髓瘤(MM)细胞系移植小鼠的成瘤情况,以及构建MM疾病模型便于体内实验研究.

方法: 利用MM细胞系LP-1、OPM2、RPMI 8226和MOLP8,以及通过转染带有Luciferase荧光素酶的慢病毒载体得到的RPMI-Luc-Puro(载体带Puromycin抗性基因)、RPMI-Luc-mCherry(载体带mCherry标记基因)和MOLP8-Luc-Puro稳定细胞系,经皮下或者尾静脉植入NOD/SCID或NSG小鼠。观察其肿瘤生长,测量肿瘤大小;应用流式细胞术检测小鼠尾血中CD138+肿瘤细胞的比例;应用血清免疫固定电泳检测外周血中的游离轻链;免疫组织化学染色法鉴定肿瘤类型;应用活体成像技术监测全身肿瘤信号.

结果: LP-1和OPM2细胞皮下植入NOD/SCID小鼠各21只,观察至7周均未见成瘤。RPMI 8226细胞皮下植入的NOD/SCID小鼠15只,1周后可观察到肿瘤形成,截至7周时,成瘤率80%(12/15)。血清免疫固定电泳可以检测到λ轻链,尾血中未检测到CD138+的肿瘤细胞;免疫组织化学染色支持浆细胞肿瘤。RPMI-Luc-Puro、RPMI-Luc-mCherry和MOLP8-Luc-Puro细胞皮下植入NOD/SCID小鼠各2只。活体成像显示RPMI-Luc-mCherry细胞植入小鼠无明显肿瘤信号,RPMI-Luc-Puro和MOLP8-Luc-Puro细胞植入小鼠1周时能探测到明显肿瘤信号,但前者2周时信号消失,后者观察至3周时肿瘤信号仍存在;这两种细胞皮下植入NSG小鼠时,两者的肿瘤信号均能持续存在。RPMI 8226、RPMI-Luc-Puro、RPMI-Luc-mCherry和MOLP8-Luc-Puro细胞经尾静脉植入NOD/SCID或NSG小鼠均未出现全身播散的肿瘤信号,仅MOLP8-Luc-Puro细胞植入的1只NSG小鼠出现过短暂的全身播散信号.

结论: 在无γ射线照射条件下,MM细胞系植入小鼠可以成瘤,有局部成瘤倾向,全身播散能力低;不同细胞系成瘤性存在较大差异,载体改造会降低细胞的成瘤能力;免疫缺陷更严重的NSG小鼠有利于肿瘤生长.

MeSH terms

  • Animals
  • Carcinogenesis
  • Cell Line
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Multiple Myeloma*