|本期目录/Table of Contents|

[1]孟荣,周杰,李晓青.骨髓间充质干细胞的分离鉴定及其对乳腺癌细胞转录组的调控作用[J].天津医科大学学报,2026,32(03):221-227.[doi:10.20135/j.issn.1006-8147.2026.03.0221]
　MENG Rong,ZHOU Jie,LI Xiaoqing.Isolation and characterization of bone marrow mesenchymal stem cells and their regulatory effects on the transcriptome of breast cancer cells[J].Journal of Tianjin Medical University,2026,32(03):221-227.[doi:10.20135/j.issn.1006-8147.2026.03.0221]
点击复制

骨髓间充质干细胞的分离鉴定及其对乳腺癌细胞转录组的调控作用(PDF)

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

卷:
32卷
期数:
2026年03期
页码:
221-227
栏目:
肿瘤疾病专题
出版日期:
2026-05-20

文章信息/Info

Title:
Isolation and characterization of bone marrow mesenchymal stem cells and their regulatory effects on the transcriptome of breast cancer cells
文章编号:
1006-8147（2026）03-0221-07
作者:
孟荣; 周杰; 李晓青
天津医科大学肿瘤医院肿瘤研究所生物化学与分子生物学研究室，国家恶性肿瘤临床医学研究中心，天津市恶性肿瘤临床研究中心，天津市肿瘤防治重点实验室，乳腺癌防治教育部重点实验室，天津300060
Author(s):
MENG Rong; ZHOU Jie; LI Xiaoqing
Department of Biochemistry and Molecular Biology， Tianjin Medical University Cancer Institute and Hospital； Tianjin Medical Univer-sity Cancer Institute&Hospital， National Clinical Research Center for Cancer； Tianjin′s Clinical Research Center for Cancer； Key Lab-oratory of Cancer Prevention and Therapy， Tianjin； Key Laboratory of Breast Cancer Prevention and Therapy， Tianjin Medical Univer-sity， Ministry of Education， Tianjin 300060， China
关键词:
骨髓间充质干细胞; 乳腺癌; 转录组分化
Keywords:
bone marrow-derived mesenchymal stem cells;  breast cancer;  transcriptome differentiation
分类号:
R73-37
DOI:
10.20135/j.issn.1006-8147.2026.03.0221
文献标志码:
A
摘要:
目的：分离并鉴定小鼠原代骨髓间充质干细胞并探讨其对MDA-MB-231乳腺癌细胞转录组的调控作用。方法：从小鼠胫骨和股骨中分离骨髓间充质干细胞，并采用3种分化诱导培养液诱导间充质干细胞向成骨、软骨、脂肪细胞分化，对分离得到的间充质干细胞的分化潜能进行鉴定。进一步收集间充质干细胞培养上清，配制条件培养液，以间接共培养的方式培养MDA-MB-231乳腺癌细胞，并采用RNA测序的方法分析间充质干细胞对乳腺癌细胞转录组的调控。结果：光学显微镜下，原代细胞呈梭形，成簇或集落式生长。间充质干细胞的成骨、成软骨、成脂三系诱导后的染色鉴定结果均为阳性。成骨诱导分化后，成骨诱导组每平方毫米细胞面积生成（124.40±2.95）个钙结节（t=42.19，P<0.001），钙结节直径为（79.85±2.58）μm（t=30.91，P<0.001），对照组没有钙结节生成；成软骨诱导组每平方毫米细胞面积有（4.20±0.41）个软骨球（t=10.09，P<0.001），直径（76.21±1.73）μm（t=44.08，P<0.001），对照组没有软骨球生成；成脂诱导组每平方毫米细胞面积有（33.80±2.18）个阳性细胞（t=15.50，P<0.001），且阳性面积百分比为57.31%±1.77%（t=32.33，P<0.001），对照组仅有微量脂滴。间充质干细胞条件培养液可以调控MDA-MB-231细胞的增殖、分化，对细胞外基质相关信号通路、转化生长因子-β超家族等信号通路也具有调控作用。结论：本研究成功分离并鉴定了小鼠原代骨髓间充质干细胞；骨髓间充质干细胞可以调控MDA-MB-231细胞的增殖和分化等生物学进程。
Abstract:
Objective： To isolate and characterize primary bone marrow-derived mesenchymal stem cells（BM-MSCs） from mice and to investigate their regulatory effects on the transcriptome of MDA-MB-231 breast cancer cells. Methods： BM-MSCs were isolated from mouse tibiae and femurs， and their multipotency was confirmed by inducing differentiation into osteoblasts， chondrocytes and adipocytes using three types of differentiation induction culture media. The differentiation potential of the isolated mesenchymal stem cells was identified. Subsequently， the culture supernatant of mesenchymal stem cells was collected， and conditioned medium was prepared. MDA-MB-231 breast cancer cells were cultured with the conditioned medium in an indirect co-culture system. The regula-tory impact of BM-MSCs on the breast cancer cell transcriptome was assessed using RNA sequencing. Results： Primary cells exhibit-ed a spindle-shaped morphology and grew in clusters or colonies under an optical microscope. Multilineage differentiation assays con-firmed the successful osteogenic， chondrogenic， and adipogenic induction of MSCs， as evidenced by positive staining. Following os-teogenic differentiation induction， the osteogenic group exhibited formation of calcium nodules（124.40±2.95） per square millime-ter of cellular area（t=42.19， P<0.001）， with an average nodule diameter of（79.85±2.58） μm（t=30.91， P<0.001）. No calcium nodules were observed in the control group. After chondrogenic induction， the chondrogenic group displayed（4.20±0.41） cartilage nod-ules per square millimeter of cellular area（t=10.09， P<0.001）， averaging diameter（76.21±1.73） μm（t=44.08， P<0.001）. The cor-responding control group showed no cartilage nodule formation. Upon adipogenic induction， the adipogenic group contained （33.80± 2.18） positively stained cells per square millimeter of cellular area（t=15.50， P<0.001）， with a positive area percentage of 57.31% ±1.77%（t=32.33， P<0.001）. The control group displayed only minimal lipid droplet formation. The conditioned medium from mesenchy-mal stem cells can regulate the proliferation and differentiation of MDA-MB-231 cells， and also exerts regulatory effects on signal-ing pathways such as those related to the extracellular matrix and the transforming growth factor-β superfamily. Conclusion： This study successfully isolates and characterizes primary BM-MSCs from mice. Furthermore， these BM-MSCs promotes biological processes in MDA-MB-231 cells， including proliferation and differentiation.

参考文献/References:

[1] SHARPE P T. Dental mesenchymal stem cells[J]. Development， 2016， 143（13）：2273-2280.
[2] SONG Z， TAO Y， JIANG R， et al. Translational potential of mes-enchymal stem cells in regenerative therapies for human diseases：challenges and opportunities[J]. Stem Cell Res Ther， 2024， 15（1）：266.
[3] MELZER C， OHE J V， HASS R. Altered tumor plasticity after dif-ferent cancer cell fusions with MSC[J]. Int J Mol Sci， 2020， 21 （21）：8347.
[4] ANKRUM J A， ONG J F， KARP J M. Mesenchymal stem cells：immune evasive， not immune privileged[J]. Nat Biotechnol， 2014， 32（3）：252-260.
[5] MELZER C， VON DER OHE J， HASS R. MSC stimulate ovarian tumor growth during intercellular communication but reduce tu-morigenicity after fusion with ovarian cancer cells[J]. Cell Commun Signal， 2018， 16（1）：67.
[6] TRIGO C M， RODRIGUES J S， CAM簟ES S P， et al. Mesenchy-mal stem cell secretome for regenerative medicine： where do we stand?[J]. J Adv Res， 2025， 70：103-124.
[7] SAMSONRAJ R M， RAGHUNATH M， NURCOMBE V， et al. Concise review： multifaceted characterization of human mesenchy-mal stem cells for use in regenerative medicine[J]. Stem Cells Transl Med， 2017， 6（12）：2173-2185.
[8] ANTOON R， OVERDEVEST N， SALEH A H， et al. Mesenchy-mal stromal cells as cancer promoters[J]. Oncogene， 2024， 43（49）：3545-3555.
[9] SHAH K. Mesenchymal stem cells engineered for cancer therapy[J]. Adv Drug Deliv Rev， 2012， 64（8）：739-748.
[10] GARCIA-APONTE O F， KAHLENBERG S， KOUROUPIS D， et al. Effects of hydrogels on mesenchymal stem/stromal cells paracrine activity and extracellular vesicles production[J]. J Extracell Vesi-cles， 2025， 14（3）：e70057.
[11] WANG J， LI R. Effects， methods and limits of the cryopreservation on mesenchymal stem cells[J]. Stem Cell Res Ther， 2024， 15（1）：337.
[12] YADAV S， MAITY P， KAPAT K. The opportunities and challenges of mesenchymal stem cells-derived exosomes in theranostics and regenerative medicine[J]. Cells， 2024， 13（23）：1956.
[13] HASS R， VON DER OHE J， UNGEFROREN H. Potential role of MSC/cancer cell fusion and EMT for breast cancer stem cell forma-tion[J]. Cancers（Basel）， 2019， 11（10）：1432.
[14] CHEN Z， XU P， WANG X， et al. MSC-NPRA loop drives fatty acid oxidation to promote stemness and chemoresistance of gastric cancer[J]. Cancer Lett， 2023， 565：216235.
[15] ZOU Z， LIN Z， WU C， et al. Micro-engineered organoid-on-a-chip based on mesenchymal stromal cells to predict immunotherapy responses of HCC patients[J]. Adv Sci（Weinh）， 2023， 10（27）：e2302640.
[16] MUNIR H， WARD L S C， MCGETTRICK H M. Mesenchymal stem cells as endogenous regulators of inflammation[J]. Adv Exp Med Biol， 2018， 1060：73-98.
[17] SHAN Y， ZHANG M， TAO E， et al. Pharmacokinetic characteris-tics of mesenchymal stem cells in translational challenges[J]. Signal Transduct Target Ther， 2024， 9（1）：242.
[18] SAXENA N， CHAKRABORTY S， DUTTA S， et al. Stiffness-de-pendent MSC homing and differentiation into CAFs-implications for breast cancer invasion[J]. J Cell Sci， 2024， 137（1）：jcs261145.
[19] JIN X， QIAO L， FAN H， et al. Long non-coding RNA MSC-AS1 facilitates the proliferation and glycolysis of gastric cancer cells by regulating PFKFB3 expression[J]. Int J Med Sci， 2021， 18（2）：546-554.
[20] HAMALIAN S， G譈TH R， RUNA F， et al. A SNAI2-PEAK1-INH-BA stromal axis drives progression and lapatinib resistance in HER2-positive breast cancer by supporting subpopulations of tumor cells positive for antiapoptotic and stress signaling markers [J]. Oncogene， 2021， 40（33）：5224-5235.
[21] RASOULI M， SAFARI F， ROUDI R， et al. Investigation of mes-enchymal stem cell secretome on breast cancer gene expression： a bioinformatic approach to identify differentially expressed genes， functional networks， and potential therapeutic targets[J]. Comput Biol Chem， 2025， 115：108331.
[22] MELZER C， OHE J V， LUO T， et al. Spontaneous fusion of MSC with breast cancer cells can generate tumor dormancy[J]. Int J Mol Sci， 2021， 22（11）：5930.
[23] LIN Z， WU Y， XU Y， et al. Mesenchymal stem cell-derived exo-somes in cancer therapy resistance： recent advances and therapeu-tic potential[J]. Mol Cancer， 2022， 21（1）：179.
[24] MU X， ZHOU Y， YU Y， et al. The roles of cancer stem cells and therapeutic implications in melanoma[J]. Front Immunol， 2024， 15：1486680.
[25] LAN T， LUO M， WEI X. Mesenchymal stem/stromal cells in cancer therapy[J]. J Hematol Oncol， 2021， 14（1）：195.
[26] GHALEH H E G， VAKILZADEH G， ZAHIRI A， et al. Investi-gating the potential of oncolytic viruses for cancer treatment via MSC delivery[J]. Cell Commun Signal， 2023， 21（1）：228.
[27] SOHRABI B， DAYERI B， ZAHEDI E， et al. Mesenchymal stem cell（MSC）-derived exosomes as novel vehicles for delivery of miR-NAs in cancer therapy[J]. Cancer Gene Ther， 2022， 29（8-9）：1105-1116.
[28] KESHTKAR S， AZARPIRA N， GHAHREMANI M H. Mesenchy-mal stem cell-derived extracellular vesicles： novel frontiers in re-generative medicine[J]. Stem Cell Res Ther， 2018， 9（1）：63.

相似文献/References:

[1]朱悦,张诗武,张丹芳,等.TA2小鼠自发乳腺癌血清蛋白质组学研究[J].天津医科大学学报,2013,19(05):373.
[2]刘 营,孙保存,刘铁菊,等.AURKA蛋白激酶在三阴乳腺癌干细胞形成血管拟态中的实验研究[J].天津医科大学学报,2013,19(06):437.
　LIU Ying,SUN Bao-cun,LIU Tie-ju,et al.Experimental study of AURKA protein kinase in the formation of vascular mimicry in triple-negative breast cancer stem cells[J].Journal of Tianjin Medical University,2013,19(03):437.
[3]伦淑敏.HOXA5基因真核表达质粒的构建及在乳腺癌细胞中的功能研究[J].天津医科大学学报,2014,20(05):337.
　LUN Shu-min. Construction of HOXA5 eukaryotic expression plasmid of and its biological significance in breast cancer cells[J].Journal of Tianjin Medical University,2014,20(03):337.
[4]伦淑敏.肌细胞增强因子2A基因真核表达质粒的构建及对乳腺癌细胞MCF-7增殖能力的影响[J].天津医科大学学报,2014,20(06):429.
　LUN Shu-min.Construction of myocyte enhancer factor 2A eukaryotic expression plasmid and effects on cell proliferation in breast cancer cell line MCF7[J].Journal of Tianjin Medical University,2014,20(03):429.
[5]孙秀梅,张 飞,田 然,等.Nanog表达上调促进乳腺癌细胞MCF-7的增殖和侵袭[J].天津医科大学学报,2014,20(06):421.
　SUN Xiu-mei,ZHANG Fei,TIAN Ran,et al.Up-regulation of Nanog promotes cell proliferation and invasion in breast cancer cells MCF-7[J].Journal of Tianjin Medical University,2014,20(03):421.
[6]张 洁,张 飞,冀 为,等. SHP2不同突变体对乳腺癌细胞的迁移和侵袭能力的影响[J].天津医科大学学报,2015,21(02):93.
　ZHANG Jie,ZHANG Fei,JI Wei,et al. Effect of different SHP2 mutants on breast cancer cell migration and invasion[J].Journal of Tianjin Medical University,2015,21(03):93.
[7]蔡 隽. FOXQ1稳定表达乳腺癌细胞系的建立及鉴定[J].天津医科大学学报,2015,21(04):292.
　CAI Jun.Establishment and identification of cell lines with stable expression of FOXQ1 in MDA-MB-231-luc[J].Journal of Tianjin Medical University,2015,21(03):292.
[8]蔡 隽 综述,冯玉梅 审校.叉头框转录因子调控乳腺癌生物学特性的研究进展[J].天津医科大学学报,2015,21(05):455.
[9]任宗娜.沉默Notch4基因对乳腺癌细胞系MDA-MB-231增殖和迁移侵袭能力的影响[J].天津医科大学学报,2015,21(06):469.
　REN Zong-na.Inhibition effect of?silencing?? Notch4 gene on the proliferation and migration and invasion activity of? breast cancer cell line?MDA-MB-231[J].Journal of Tianjin Medical University,2015,21(03):469.
[10]周岩,宋伟杰,张飞,等.人附睾蛋白4在乳腺癌发生发展中的机制研究[J].天津医科大学学报,2015,21(06):466.
　ZHOU Yan,SONG Wei-jie,ZHANG Fei,et al.Mechanism of human epididymis protein 4 in development and progression of breast cancer[J].Journal of Tianjin Medical University,2015,21(03):466.

备注/Memo

备注/Memo:
基金项目 天津市医学重点学科建设资助（TJYXZDXK-3-003A）
作者简介 孟荣（2000-），女，硕士在读，研究方向：肿瘤生物化学与分子生物学；通信作者：李晓青，E-mail：xqli@tmu.edu.cn。
（2025-10-21收稿）
更新日期/Last Update: 2026-05-25