Construction of the Bone Metastasis Model in B6-hRANKL Mice and Exploration of the Efficacy of hRANKL Monoclonal Antibody

doi:10.19586/j.2095-2341.2025.0098

Current Biotechnology ›› 2025, Vol. 15 ›› Issue (6): 1077-1085.DOI: 10.19586/j.2095-2341.2025.0098

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Construction of the Bone Metastasis Model in B6-hRANKL Mice and Exploration of the Efficacy of hRANKL Monoclonal Antibody

Lili BI¹^,²^,³(), Yunxia ZHAO¹^,²^,³(), Kui JIANG¹^,²^,³, Yuanyuan WANG¹^,²^,³, Ruiping LI¹^,²^,³

^1.Hualan Genetic Engineering Co. ，Ltd. ，Henan Xinxiang 453500，China
^2.Henan Biomedical Industry Research Institute，Henan Xinxiang 453200，China
^3.Henan Provincial Key Laboratory of Recombinant Protein Drugs，Henan Xinxiang 453200，China

Received:2025-08-01 Accepted:2025-09-05 Online:2025-11-25 Published:2026-01-04
Contact: Yunxia ZHAO

Abstract

Abstract:

To evaluate the pharmacological efficacy of hRANKL monoclonal antibody and explore the underlying pharmacodynamic mechanisms of tumor bone metastasis， the B6-hRANKL mouse model of osteolytic bone metastasis was established by injecting LLC1-luc into the tibial bone marrow cavity. LLC1-luc cells were injected into the tibial bone marrow cavity of B6-hRANKL mice. The model efficacy was evaluated through mouse live imaging， bone mineral density X-ray imaging， tartrate-resistant acid phosphatase 5b （TRACP 5b） detection， and histological analysis. After model establishment， eligible mice were randomly divided into 0.9% sodium chloride control group and the drug administration group. On the basis of the model evaluation indicators， immunohistochemistry was used to verify the distribution of the drug in bone tissue. The results showed that the osteolytic bone metastasis model（B6-hRANKL-LLC1-luc model）of tumor was successfully established， exhibiting significant bone density reduction （BMD decrease in LLC1-luc cell group vs. physiological saline group， P0.05）， elevated TRACP5b activity （P0.05）， and tumor cell bone infiltration. Although the hRANKL monoclonal antibody treatment group demonstrated drug distribution in femur and tibia bones， it failed to improve bone destruction markers. The above results suggestted that the B6-hRANKL-LLC1-luc model established in this study can be utilized for investigating osteolytic bone metastasis mechanisms. However， hRANKL monoclonal antibody did not demonstrate the expected efficacy in this model， which may be attributed to the biological characteristics of the tumor， mechanism of action compatibility issues， and inherent limitations of the model itself.

Key words: B6-hRANKL mice, LLC1-luc cells, osteolytic bone metastasis model, hRANKL monoclonal antibody

摘要：

研究旨在建立B6-hRANKL小鼠胫骨骨髓腔注射LLC1-luc溶骨性骨转移模型，评价hRANKL单抗的药效作用，探讨肿瘤骨转移的潜在药效作用机制。在B6-hRANKL小鼠胫骨骨髓腔注射LLC1-luc细胞，通过小鼠活体成像、骨密度X射线成像、抗酒石酸酸性磷酸酶5b（TRACP 5b）检测及组织学分析评估模型有效性；模型构建后，可入组小鼠随机分为0.9%氯化钠对照组和给药组，在模型评价指标基础上，采用免疫组化法验证药物骨组织分布。实验结果表明，肿瘤溶骨性骨转移模型（B6-hRANKL-LLC1-luc模型）构建成功，表现为骨密度（bone mineral density， BMD）降低（LLC1-luc细胞组BMD较0.9%氯化钠组显著下降，P0.05）、TRACP5b显著升高（P0.05）及肿瘤细胞骨浸润。hRANKL单抗治疗组虽显示在小鼠股骨和胫骨有药物分布，但未改善骨破坏指标。实验结果表明，B6-hRANKL-LLC1-luc模型可用于溶骨性骨转移机制研究，但hRANKL单抗在此模型中未表现出预期疗效，可能与肿瘤生物学特性限制、药物作用机制适配性及模型局限性有关。

关键词: B6-hRANKL小鼠, LLC1-luc细胞, 骨转移模型, hRANKL单抗

CLC Number:

Q511

Lili BI, Yunxia ZHAO, Kui JIANG, Yuanyuan WANG, Ruiping LI. Construction of the Bone Metastasis Model in B6-hRANKL Mice and Exploration of the Efficacy of hRANKL Monoclonal Antibody[J]. Current Biotechnology, 2025, 15(6): 1077-1085.

毕利利, 赵云霞, 江魁, 王园园, 李瑞萍. B6-hRANKL小鼠的骨转移模型构建及hRANKL单抗疗效探讨[J]. 生物技术进展, 2025, 15(6): 1077-1085.

Figures/Tables 15

Fig. 1 Trend of body weight changes in modeling mices

Fig. 2 In vivo imaging of individual mice in different groups

Fig. 3 Changes of photon numbers in live imaging of mices in different groups

Fig. 4 Changes in bone density of mices in different groups

Fig. 5 Example image of HE staining of femoral tissue

Fig. 6 Example image of HE staining of tibial tissue

Fig. 7 TRACP 5b levels in the serum of mices from different groups

Fig. 8 Live imaging of individual mice in different groups after administration

Fig. 9 Changes of photon numbers in live imaging of mices in different groups after administration

Fig. 10 Changes in bone density of mices in different groups after administration

Fig. 11 TRACP 5b levels in the serum of mices from different groups after administration

Fig. 12 Example image of HE staining of femoral tissue after administration

Fig. 13 Example image of HE staining of tibial tissue after administration

Fig. 14 Distribution of the drug in the femur of mice(20×)

Fig. 15 Distribution of the drug in the tibia of mice(20×)

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Construction of the Bone Metastasis Model in B6-hRANKL Mice and Exploration of the Efficacy of hRANKL Monoclonal Antibody

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