Page 7 - 南京医科大学学报自然科学版

第43卷第1期 南京医科大学学报（自然科学版）
2023年1月 Journal of Nanjing Medical University（Natural Sciences） · 1 ·

·基础研究·

多西环素通过 MEK/ERK 信号通路促进 MC3T3⁃E1 细胞株体外

成骨分化

耿佳伟 ，费小明 ，汤 郁 ，李海璐 ，王丽霞 1
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2
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1
江苏大学附属医院血液科，风湿科，江苏 镇江 212000
1 2
［摘 要］ 目的：研究在体外条件下多西环素（doxycycline，DOX）对前成骨细胞株 MC3T3⁃E1 成骨分化的影响及可能机制。
方法：在成骨诱导剂体外诱导MC3T3⁃E1细胞株成骨分化条件下，茜素红染色检测成骨细胞分化；实时定量PCR检测DOX对
MC3T3⁃E1细胞相关成骨基因OCN、Runx2的影响；用DOX、MEK抑制剂（U0126）单独或联合处理MC3T3⁃E1细胞后，Western blot
检测N⁃cadherin、p⁃MEK及p⁃ERK等蛋白的表达。结果：在MC3T3⁃E1细胞株体外成骨诱导分化中，加入DOX可以增强茜素红染
色阳性率。DOX上调MC3T3⁃E1细胞相关成骨基因OCN、Runx2的表达。DOX处理MC3T3⁃E1细胞后，其N⁃cadherin蛋白表达水
平下降（P < 0.05），p⁃MEK和p⁃ERK蛋白的表达增加（P < 0.05）。而MEK拮抗剂（U0126）则显著上调N⁃cadherin蛋白表达水平，
同时降低p⁃MEK、p⁃ERK水平。用U0126联合DOX处理MC3T3⁃E1细胞后，DOX对N⁃cadherin、p⁃MEK和p⁃ERK蛋白水平的影响
可被U0126拮抗。在MC3T3⁃E1细胞株体外成骨诱导分化中，同时存在DOX和U0126时，茜素红染色阳性率低于单独DOX组。
结论：DOX可以促进MC3T3⁃E1细胞株的体外成骨分化；而DOX的这一效应可能是通过MEK⁃ERK信号通路参与完成的。
［关键词］ 多发性骨髓瘤；多西环素；成骨分化；MC3T3⁃E1；N⁃cadherin；MEK/ERK信号通路
［中图分类号］ R733.3 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）01⁃001⁃08
doi：10.7655/NYDXBNS20230101

Doxycycline enhances in vitro osteogenic differentiation of MC3T3⁃E1 cells via MEK/ERK

signaling pathway
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GENG Jiawei ，FEI Xiaoming ，TANG Yu ，LI Hailu ，WANG Lixia 1
Department of Hematology，Department of Rheumatology，the Affiliated Hospital of Jiangsu University，Zhenjiang
1 2
212000，China
［Abstract］ Objective：To investigate the effects of doxycycline on in vitro osteogenic differentiation of pro ⁃ osteoblasts cell line
MC3T3⁃E1 and the possible mechanisms. Methods：Alizarin red staining was employed to detect the osteogenic differentiation. Real⁃
time PCR was used to detect the effect of DOX on the expression of OCN and Runx2 in MC3T3⁃E1 cells. MC3T3⁃E1 cells were treated
with either DOX，MEK inhibitor U0126 alone or in combination，and the levels of N⁃cadherin，p⁃MEK and p⁃ERK were detected by
Western blot. Results：When DOX was added during osteogenic induction，alizarin red staining positivity of the cultured MC3T3⁃E1
was significantly enhanced. DOX increased the expression of OCN and Runx2 in MC3T3⁃E1 cells. Besides，DOX decreased the levels
of N⁃cadherin and increased the level of p⁃MEK and p⁃ERK in MC3T3⁃E1（P < 0.05）. On the contrary，MEK antagonist U0126
significantly increased the expression of N⁃cadherin protein and decreased the levels of p⁃MEK and p⁃ERK（P < 0.05）. When MC3T3⁃E1
cells were treated with DOX in the presence of MEK inhibitor U0126，the changes in N⁃cadherin，p⁃MEK and p⁃ERK were showed to
be reversed comparing with DOX alone. When both U0126 and DOX were present during in vitro osteogenic differentiation of MC3T3⁃E1，
alizarin red staining positivity was less observed than that of DOX alone. Conclusion：In this study，DOX is showed to enhance the
in vitro osteogenic differentiation of MC3T3⁃E1，which is probably associated with MEK/ERK signaling pathway.
［Key words］ multiple myeloma；doxycycline；osteogenesis；MC3T3⁃E1；N⁃cadherin；MEK/ERK signaling pathway
［J Nanjing Med Univ，2023，43（01）：001⁃007，106］

［基金项目］ 江苏省卫生健康委科研课题（H2018084）；江苏省社会发展重点项目（临床前沿技术）课题（BE202068）
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通信作者（Corresponding author），E⁃mail：feixiaomingujs@aliyun.com