成骨前体细胞来源的外泌体对乳腺癌细胞迁移和增殖影响的研究

阳琴娜; 元欣欣; 杜锐凯; 厉建伟; 李英贤; 千年松

doi:10.3969/j.issn.2095-5227.2023.04.012

成骨前体细胞来源的外泌体对乳腺癌细胞迁移和增殖影响的研究

doi: 10.3969/j.issn.2095-5227.2023.04.012

阳琴娜^{1, 2,},
元欣欣²,
杜锐凯²,
厉建伟²,
李英贤²,
千年松^3, ,

1.
解放军医学院，北京　100853
2.
中国航天员科研训练中心航天医学基础与应用国家重点实验室，北京　100094
3.
解放军总医院第八医学中心呼吸与危重症医学部胸部肿瘤科，北京　100039

基金项目: 军委后勤保障部卫生局军队医学科技青年培育计划拔尖项目(2020NPY112)；海南省重点研发项目(ZDYF2023SHFZ117)

详细信息

作者简介:
阳琴娜，女，在读硕士。Email: cheernanayang@163.com

通讯作者:
千年松，博士，副主任医师，副教授，科室主任。Email: 18701317301@qq.com

中图分类号: R329.2
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- 被引次数: 0
出版历程
- 收稿日期: 2022-06-24
- 网络出版日期: 2023-04-03
- 刊出日期: 2023-04-28

Effects of osteogenic precursor cells derived exosomes on migration and proliferation of breast cancer cells

1.
Chinese PLA Medical School, Beijing 100853, China
2.
State Key Lab of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing 100094, China
3.
Department of thoracic Oncology, Senior Department of Respiratory and Critical Care Medicine, the Eighth Medical Center, Chinese PLA General Hospital, Beijing 100039, China

More Information

Corresponding author: QIAN Niansong. Email: 187013173013@qq.com

摘要

摘要: 背景乳腺癌骨转移是目前研究热点之一，骨微环境中成骨细胞可以促进肿瘤细胞存活并形成微转移，但目前鲜有成骨细胞来源的外泌体对乳腺癌作用的研究报道。目的探讨成骨前体细胞MC3T3-E1来源的外泌体对乳腺癌4T1和MDA-MB-231细胞迁移和增殖的影响。方法通过超高速离心法提取MC3T3-E1细胞来源的外泌体，透射电子显微镜、纳米颗粒跟踪分析(nanopartic tracking analysis，NTA)和Western blot方法鉴定外泌体特性；利用划痕实验和Transwell实验检测分析MC3T3-E1来源的外泌体对乳腺癌4T1和MDA-MB-231细胞迁移的影响；CCK-8法和克隆形成实验检测MC3T3-E1来源的外泌体对乳腺癌4T1和MDA-MB-231细胞增殖的影响。结果 MC3T3-E1细胞来源的外泌体具有膜结构囊状小泡，直径分布为100 ~ 200 nm，且表达外泌体标志蛋白Alix和TSG101。CCK-8活性实验结果显示，浓度为20 μg/mL的MC3T3-E1细胞来源的外泌体不影响乳腺癌细胞的活性，30 μg/mL、50 μg/mL的外泌体能够降低乳腺癌细胞活性。划痕实验和Transwell实验结果显示，20 μg/mL外泌体能够促进乳腺癌细胞的迁移(P＜0.05)。CCK-8增殖实验和细胞克隆形成实验结果显示，20 μg/mL外泌体能够促进乳腺癌细胞的增殖(P＜0.05)。结论 MC3T3-E1来源的外泌体可以促进乳腺癌细胞的迁移和增殖，这为乳腺癌骨转移的治疗提供可能的靶点和治疗策略。
- 成骨前体细胞MC3T3-E1 /
- 外泌体 /
- 乳腺癌 /
- 迁移 /
- 增殖
Abstract: Background Bone metastasis of breast cancer is one of the current research hotspots. Osteoblasts in the bone microenvironment can promote the survival of tumor cells and form micrometastasis. While, there are few studies on the effect of osteoblast derived exosomes on breast cancer. Objective To investigate the effects of MC3T3-E1 exosomes on the migration and proliferation of breast cancer 4T1 and MDA-MB-231 cells. Methods Exosomes derived from MC3T3-E1 cells were extracted, transmission electron microscopy (TEM), nanopartic tracking analysis (NANOPartic tracking), NTA and Western blot were used to identify exosomes. Transwell and scratch assay were used to detect the effect of MC3T3-E1-derived exosomes on the migration of breast cancer 4T1 and MDA-MB-231 cells. The effects of MC3T3-E1 derived exosomes on the proliferation of breast cancer cells were detected by CCK-8 assay and clonogenesis assay. Results The exosomes derived from MC3T3-E1 cells were characterized by membrane-structured cystic vesicles under transmission electron microscopy. The exosomes derived from MC3T3-E1 cells were 100-200 nm in diameter and expressed exosome related proteins Alix and TSG101. CCK-8 activity test results showed that exosomes derived from MC3T3-E1 cells at a concentration of 20 μg/mL did not affect the vitality of breast cancer cells, while exosomes derived from 30 μg/mL and 50 μg/mL could reduce the vitality of breast cancer cells. Scratch test and Transwell test results showed that 20 μg/mL of exosomes could promote the migration of breast cancer cells (P<0.05). CCk-8 proliferation assay and colony formation assay showed that 20 μg/mL exosomes could promote the proliferation of breast cancer cells (P<0.05). Conclusion Exosomes derived from MC3T3-E1 can promote the migration and proliferation of breast cancer cells, which provides a potential target for therapeutic strategy on bone metastasis of breast cancer.
- osteogenic precursor cell MC3T3-E1 /
- exosome /
- breast cancer /
- migration /
- proliferation

HTML全文

图 1 外泌体纯度鉴定

A：透射电子显微镜下结果；B：纳米颗粒跟踪分析结果；C：外泌体相关蛋白鉴定；D：外泌体相关蛋白灰度分析

Figure 1. Identification of exosomes

A: TEM image; B:NTA results; C: Identification of exosomes related proteins; D: Gray level analysis of exosomes related proteins

下载: 全尺寸图片幻灯片

图 2 MC3T3-E1细胞来源外泌体对乳腺癌细胞活性的影响

A：4T1细胞CCK-8活性实验定量结果；B：MDA-MB-231细胞CCK-8活性实验定量结果

Figure 2. MC3T3-E1 derived exosomes on the viability of breast cancer cells

A: Quantitative results of CCK-8 activity assay in 4T1 cells; B: Quantitative results of CCK-8 activity assay in MDA-MB-231 cells

下载: 全尺寸图片幻灯片

图 3 MC3T3-E1细胞来源外泌体对乳腺癌细胞迁移能力的影响(200×)

A：划痕实验镜下观察和定量结果；B：Transwell实验镜下观察和定量结果

Figure 3. MC3T3-E1 derived exosomes on the migration of breast cancer cells (200×)

A: Observation under the microscope of the scratch test and quantitative results of the scratch test; B: Observation under the microscope of the transwell test and quantitative results of the transwell test

下载: 全尺寸图片幻灯片

图 4 MC3T3-E1细胞来源外泌体对乳腺癌细胞增殖能力的影响

A：CCK-8增殖实验定量结果；B：克隆形成实验和定量结果

Figure 4. Effecf on MC3T3-E1 derived exosomes on the proliferation of breast cancer cells

A: Quantitative results of CCK-8 proliferation experiment; B: Colony formation assay and quantitative results

下载: 全尺寸图片幻灯片

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