Page 46 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第41卷第7期
·976 · Journal of Nanjing Medical University（Natural Sciences） 2021年7月

·基础医学·

GDF15激活ERK/Bcl⁃2通路增强BM⁃MSCs放射抗性

吴 春 ，张小梅 ，向 阳 ，冉 茜 ，李忠俊 ，张克斌 2*
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陆军军医大学第二附属医院检验医学中心（输血科）放射生物学实验室，临床医学研究中心，重庆 400037
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［摘 要］ 目的：探讨生长分化因子15（growth differentiation factor 15，GDF15）在调控人骨髓间充质干细胞（bone marrow mes⁃
enchymal stem cells，BM⁃MSCs）放射抗性中的作用及机制。方法：利用Lipofectamine 3000转染siRNA，干扰BM⁃MSCs的GDF15
表达。9 Gy Co⁃60 γ射线照射后，采用流式细胞术检测细胞周期及凋亡，CCK⁃8检测增殖，JC⁃1检测线粒体膜电位，免疫荧光、
实时荧光定量PCR（RT⁃qPCR）和蛋白免疫印迹分析（Western blot）检测GDF15及ERK/Bcl⁃2信号通路关键分子的表达。结果：
放射后GDF15的表达水平显著升高。未放射条件下，干扰GDF15对细胞周期、增殖和凋亡无显著影响（P ˃ 0.05）。放射后，干
扰GDF15导致细胞G2期阻滞增加（P ˂ 0.05），细胞增殖显著抑制（P ˂ 0.01）；同时线粒体膜电位降低，Caspase3活化及细胞凋
亡增加（P ˂ 0.05）。干扰GDF15显著抑制ERK1/2的磷酸化和Bcl⁃2的蛋白水平，而增加Bax的表达。结论：放射诱导GDF15上
调，可提高BM⁃MSCs的放射抗性，其机制与ERK/Bcl⁃2信号通路有关。
［关键词］ 骨髓间充质干细胞；GDF15；放射抗性；凋亡；增殖
［中图分类号］ R818 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）07⁃976⁃09
doi：10.7655/NYDXBNS20210707

GDF15 promotes irradiation resistance of bone marrow mesenchymal stem cells via the
activation of ERK/Bcl⁃2 pathway
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WU Chun ，ZHANG Xiaomei ，XIANG Yang ，RAN Qian ，LI Zhongjun ，ZHANG Kebin 2*
Laboratory Medicine Center，Department of Blood Transfusion，Lab of Radiation Biology，Clinical Medical Research
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Center，the Second Affiliated Hospital，Third Military Medical University，Chongqing 400037，China
［Abstract］ Objective： To investigate the role of growth differentiation factor 15（GDF15）in irradiation resistance of human bone
marrow mesenchymal stem cells（BM⁃MSCs）and determine its underlying mechanism. Methods：The BM⁃MSCs were transfected with
GDF15 siRNA using Lipofectamine 3000. After the irradiation of BM⁃MSCs with 9 Gy Co⁃60 γ⁃rays，the cell cycle and apoptosis were
detected by flow cytometry，CCK⁃8 was used to detect cell proliferation，and JC⁃1 was used to detect mitochondrial membrane potential，
immunofluorescence，quantitative real⁃time PCR（RT⁃qPCR）and Western blot were used to detect the expression of GDF15，and the
key molecules in ERK/Bcl ⁃ 2 signaling pathway. Results：After irradiation，the expression of GDF15 was increased significantly.
Interference of GDF15 had no significant effect on cell cycle，proliferation and apoptosis in no ⁃ irradiated BM ⁃ MSCs（P ˃ 0.05）.
Interference of GDF15 in BM ⁃ MSCs suffering form irradiation increased G2 phase arrest（P ˂ 0.05），inhibited cell proliferation
significantly（P ˂ 0.01），and decreased mitochondrial membrane potential. Meanwhile，Caspase3 was activated and apoptosis was
significantly increased（P ˂ 0.05）. The phosphorylation level of ERK1/2 and the expression level of Bcl⁃2 in GDF15 interference cells
decreased significantly，while the expression of Bax increased significantly. Conclusion：The up ⁃ regulated expression of GDF15 ⁃
induces upon irradiation，and enhances irradiation resistance of BM⁃MSCs through the activation of ERK/Bcl⁃2 signaling pathway.
［Key words］ bone marrow mesenchymal stem cells；GDF15；irradiation resistance；apoptosis；proliferation
［J Nanjing Med Univ，2021，41（07）：976⁃983，991］

［基金项目］ 中国博士后科学基金（2018M643861）；陆军军医大学第二附属医院人才培养计划（2018YQYLY005）
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通信作者（Corresponding author），E⁃mail： zhongjunli@tmmu.edu.cn；zhangkebin12@163.com