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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

24卷

期数:

2018年03期

页码:

205-208

栏目:

出版日期:

2018-05-20

文章信息/Info

Title:

Study on the method of inducing osteoclast maturation with breast cancer cell MDA-MB-231

作者:

王硕

天津医科大学肿瘤医院生物化学与分子生物学研究室，国家肿瘤临床医学研究中心，天津市“肿瘤防治”重点实验室，天津市恶性肿瘤临床医学研究中心，乳腺癌防治教育部重点实验室，天津 300060

Author(s):

WANG Shuo

Department of Biochemistry and Molecular Biology，Cancer Institute and Hospital，Tianjin Medical University, National Clinical Research Center for Cancer，Key Laboratory of Cancer Prevention and Therapy, Tianjin，Tianjin’s Clinical Research Center for Cancer，Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education，Tianjin 300060, China

关键词:

M-CSF; RANKL; 破骨细胞; 乳腺癌; 炎症反应

Keywords:

M-CSF;  RANKL;  osteoclast;  breast cancer;  inflammatory reaction

分类号:

R737.9

DOI:

-

文献标志码:

A

摘要:

目的：探索乳腺癌细胞诱导小鼠骨髓源性破骨细胞成熟最佳条件。方法：采用贴壁分选法和贴壁前加入15 ng/mL巨噬细胞集落刺激因子（M-CSF）贴壁分选法分离得到小鼠骨髓单核细胞。两种分选方法得到的小鼠骨髓单核细胞用30 ng/mL M-CSF处理3 d后，分别均分为两组，一组用50 ng/mL核因子κB受体活化因子配体（RANKL），30 ng/mL M-CSF和20% MDA-MB-231细胞上清处理7 d；另一组用50 ng/mL RANKL和30 ng/mL M-CSF处理2 d后再用50 ng/mL RANKL，30 ng/mL M-CSF和20% MDA-MB-231细胞上清处理7 d，TRAP染色，统计分析TRAP阳性细胞数和直径大小。结果：50 ng/mL RANKL，30 ng/mL M-CSF和20% MDA-MB-231细胞条件培养基诱导，细胞发生炎症反应；50 ng/mL RANKL和30 ng/ml M-CSF处理2 d后，50 ng/mL RANKL，30 ng/mL M-CSF和20% MDA-MB-231细胞条件培养基能够诱导出成熟的破骨细胞；相比直接贴壁分选，贴壁前加入15 ng/mL M-CSF贴壁分选能够分选出更多具有活力的前体破骨细胞。结论：诱导破骨细胞分化最佳条件为：贴壁前加入15 ng/mL M-CSF处理24 h，30 ng/mL M-CSF 处理3 d后，用50 ng/mL RANKL和30 ng/mL M-CSF处理2 d，再用50 ng/mL RANKL，30 ng/mL M-CSF和20% MDA-MB-231细胞条件培养基诱导7 d。

Abstract:

Objective: To investigate the optimal conditions for the maturation of mouse bone marrow-derived osteoclast induced by breast cancer cells. Methods: Bone marrow mononuclear cells were obtained by adherence and the method that the cells were treated with 15 ng/mL macrophage colony stimulating factor (M-CSF) before adherence. After treatment with 30 ng/mL M-CSF for 3 days, murine bone marrow mononuclear cells were divided into two groups. One group was treated with 50 ng/mL receptor activator of nuclear factor kappa B ligand (RANKL), 30 ng/mL M-CSF and 20% MDA-MB-231 cell conditioned medium for 7 days, and the other group of which was treated with 50 ng/mL RANKL and 30 ng/mL M-CSF for 2 days first and, subsequently, treated with 50 ng/mL RANKL, 30 ng/mL M-CSF and 20% MDA-MB-231 cell conditioned medium for 7 days. TRAP staining and statistical analysis were carried out. Results: The results showed that inflammation was observed in the cells induced by 50 ng/mL RANKL, 30 ng/mL M-CSF and 20% MDA-MB-231 cell conditioned medium; however, mature osteoclasts was induced after treatment with 50 ng/mL RANKL and 30 ng/mL M-CSF for 2 days; And, 15 ng/mL M-CSF was essential before the adherence to sort out more viable precursor osteoclast cells, compared with the direct cell adherence method. Conclusion: The optimal conditions are as following: the cells should be treated with 15 ng/mL M-CSF before adherence for 24 h. After treated with 30 ng/mL M-CSF for 3 days, the cells should be treated with 50 ng/mL RANKL and 30 ng/mL M-CSF for 2 days. And 20% MDA-MB-231 conditioned medium ought to be added for 7 days.

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相似文献/References:

[1]赵立明综述,胡永成审校.骨巨细胞瘤多学科联合的诊治进展[J].天津医科大学学报,2016,22(01):90.

备注/Memo

备注/Memo:

文章编号 1006-8147(2018)03-0205-04
作者简介 王硕（1993-），男，硕士在读，研究方向：生物化学与分子生物学，E-mail：1018579850@qq.com。

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更新日期/Last Update: 2018-05-20