中国组织工程研究Issue(50)：8048-8055,8.DOI:10.3969/j.issn.2095-4344.2015.50.003
let-7f对骨髓间充质干细胞增殖的影响
let-7f effects on the proliferation of bone marrow mesenchymal stem cells
摘要
Abstract
BACKGROUND:There is no clear understanding about the effect of let-7f and interleukin-6 (IL-6) on the proliferation of bone marrow mesenchymal stem cels and their relationship. <br> OBJECTIVE: To explore the effects of expression levels of let-7f and IL-6 on the proliferation of bone marrow mesenchymal stem cels and their relationship. <br> METHODS:(1) LV-rno-let-7f-up and LV-rno-let-7f-down were constructed and transfected into bone marrow mesenchymal stem cels of Sprague-Dawley rats, respectively. Then, there were four groups in the study: transfection upregulation group transfected with LV-rno-let-7f-up), transfection inhibition group (transfected with LV-rno-let-7f-down), negative control group (transfected with FU-RNAi-NC-LV), and untransfected group. The expression level of let-7f in each group was detected by qRT-PCR. The proliferation ability of cels and expression levels of IL-6 when let-7f expression was at different levels were detected by MTT, flow cytometry and ELISA. The expression of Cyclin D1 at mRNA and protein levels was detected by qRT-PCR and western blot, respectively. (2) To predict the potential target gene of let-7f, the wild-type/mutant IL-6 3’UTR reporter gene vectors were constructed, and cotransfected with let-7f/let-7f inhibitor respectively into the 293T cels to measure the luciferase. <br> RESULTS AND CONCLUSION: Compared with the negative control group, the proliferative and cloning capacities of cels in the transfection upregulation group were higher; the number of cels was significantly decreased at G1 stage and increased at S stage, and the apoptotic cels were reduced in number (P < 0.05). However, the transfection inhibition group had opposite results. The expression level of IL-6 in the transfection upregulation group was lower than that in the untransfected group and negative control group (P < 0.05); while in the transfection inhibition group, the expression level of IL-6 was significantly increased (P < 0.05). The expression of Cyclin D1 at mRNA and protein levels was up-regulated in transfection upregulation group (P < 0.05) and down-regulated in the transfection inhibition group (P < 0.05), but there was no significant difference between the negative control group and untransfected group (P > 0.05). Luciferase activity of cels transfected with wide-type IL-6 3’UTR and let-7f was significantly reduced (P < 0.05). These findings indicate that up-regulation of let-7f can promote the proliferative and cloning capacities of bone marrow mesenchymal stem cels and reduce cel apoptosis, but downrelation of let-7f exhibits an inhibitory effect. Overexpression of IL-6 can suppress the proliferation of bone marrow mesenchymal stem cels, which is considered to be a target gene of let-7f, and let-7f may suppress the expression of IL-6 to promote the cel proliferation.关键词
干细胞/骨髓干细胞/细胞增殖/let-7f/骨髓间充质干细胞/慢病毒/白细胞介素6/国家自然科学基金Key words
Bone Marrow/Mesenchymal Stem Cels/Interleukin-6/Cel Proliferation/Tissue Engineering分类
医药卫生引用本文复制引用
李明哲,龚哲,王静,王以文,姚要兵,侯晓灿,焦淑洁,滕军放,贾延劼..let-7f对骨髓间充质干细胞增殖的影响[J].中国组织工程研究,2015,(50):8048-8055,8.基金项目
国家自然科学基金项目骨髓间质干细胞神经命运决定中的自噬现象及其mTOR 信号通路的作用(81071114)；Lin28/Let-7调控的骨髓间充质干细胞移植治疗阿尔茨海默病的实验研究(81371385)Funding：the National Natural Science Foundation of China, No.81071114,81371385 （81071114）
