[1]方 宇,董重阳,高斌礼,等.微小RNA-214靶向调控PTEN对成骨细胞MC3T3-E1增殖、分化、矿化的影响及机制研究[J].陕西医学杂志,2023,52(12):1619-1623,1630.[doi:DOI:10.3969/j.issn.1000-7377.2023.12.001]
　FANG Yu,DONG Chongyang,GAO Binli,et al.Effects of miR-214 on proliferation,differentiation and mineralization of osteoblast MC3T3-E1 by targeting PTEN and its mechanism[J].,2023,52(12):1619-1623,1630.[doi:DOI:10.3969/j.issn.1000-7377.2023.12.001]
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微小RNA-214靶向调控PTEN对成骨细胞MC3T3-E1增殖、分化、矿化的影响及机制研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
52
期数:
2023年12期
页码:
1619-1623,1630
栏目:
基础研究
出版日期:
2023-12-05

文章信息/Info

Title:
Effects of miR-214 on proliferation,differentiation and mineralization of osteoblast MC3T3-E1 by targeting PTEN and its mechanism
作者:
方 宇1; 董重阳2; 高斌礼1; 李明宇1; 李 伟3; 郭 文1; 焦志超1
(1.内蒙古医科大学附属医院骨科,内蒙古 呼和浩特 010050; 2.内蒙古医科大学中医学院,内蒙古 呼和浩特 010107; 3.内蒙古医科大学第二附属医院影像中心,内蒙古 呼和浩特 010010)
Author(s):
FANG Yu; DONG Chongyang; GAO Binli; LI Mingyu; LI Wei; GUO Wen; JIAO Zhichao
(Department of Orthopaedics,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China)
关键词:
骨质疏松;  微小RNA-214;  第10号染色体丢失性磷酸酶及张力蛋白同源基因;  磷酸化磷脂酰肌醇3-激酶/蛋白激酶B信号通路;  成骨细胞MC3T3-E1;  增殖;  分化;  矿化
Keywords:
Osteporsis;  miR-214;  PTEN;  PI3K/AKT signaling pathway;  Osteoblasts MC3T3-E1 cell;  Proliferation;  Differentiation;  Mineralization
分类号:
R 580
DOI:
DOI:10.3969/j.issn.1000-7377.2023.12.001
文献标志码:
A
摘要:
目的:探讨微小RNA-214(miR-214)靶向调控第10号染色体丢失性磷酸酶及张力蛋白同源基因(PTEN)对成骨细胞MC3T3-E1增殖、分化、矿化的影响及机制。方法:将成骨细胞MC3T3-E1分为miR-124 mimic组(转染miR-124 mimic)、miR-124 NC组(转染miR-124 NC)、pcDNA3.1-PTEN组(转染pcDNA3.1-PTEN)、miR-124 mimic+pcDNA3.1-PTEN组(转染miR-124 mimic+pcDNA3.1-PTEN)。采用在线生信预测miR-124和PTEN靶向结合位点,通过双荧光素酶报告基因进行验证。采用实时荧光定量聚合酶链式反应(RT-qPCR)检测细胞miR-124和PTEN mRNA表达。采用CCK-8法检测细胞增殖。采用碱性磷酸酶活性测定检测细胞分化能力。采用茜素红染色和骨钙素水平测定检测细胞的矿化。采用Werstern blot检测PTEN、Ⅰ型胶原蛋白(Col Ⅰ)、骨桥蛋白(OPN)、磷酸化磷脂酰肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)水平。结果:与miR-124 NC组比较,miR-124 mimic组野生型PTEN的荧光素酶活性降低(均P<0.05)。与miR-124 NC组比较,miR-124 mimic组miR-124及p-PI3K、p-AKT蛋白表达升高,PTEN mRNA和蛋白、细胞增殖率、碱性磷酸酶活性、Col Ⅰ和OPN蛋白、细胞矿化率和骨钙素水平降低,而pcDNA3.1-PTEN组则相反(均P<0.05)。与miR-124 mimic组比较,miR-124 mimic+pcDNA3.1-PTEN组p-PI3K、p-AKT蛋白表达降低,PTEN mRNA和蛋白、细胞增殖率、碱性磷酸酶活性、Col Ⅰ和OPN蛋白、细胞矿化率和骨钙素水平升高(均P<0.05)。结论:miR-214通过靶向下调PTEN蛋白表达激活PI3K/AKT信号通路,从而调控成骨细胞MC3T3-E1增殖、分化和矿化。
Abstract:
Objective:To investigate the effects of miR-214 on proliferation,differentiation and mineralization of osteoblast MC3T3-E1 by targeting PTEN and its mechanism.Methods:MC3T3-E1 cells were divided into miR-124 mimic group(transfected with miR-124 mimic),miR-124 NC group(transfected with miR-124 NC),pcDNA3.1-PTEN group(transfected with pcDNA3.1-PTEN)and miR-124 mimic+pcDNA3.1-PTEN group(transfected with miR-124 mimic+pcDNA3.1-PTEN).The target binding sites of miR-124 and PTEN were predicted by online bioinformatics and verified by dual luciferase reporter gene.The expression of miR-124 and PTEN mRNA was detected by RT-qPCR.Cell proliferation was detected by CCK-8 assay.Alkaline phosphatase activity assay was used to detect the differentiation ability of cells.The mineralization of the cells was detected by alizarin red staining and osteocalcin level determination.The levels of PTEN,Col Ⅰ,OPN,p-PI3K,and p-AKT were detected by Werstern blot.Results:Compared with miR-124 NC group,the luciferase activity of wild-type PTEN in miR-124 mimic group decreased(all P<0.05).Compared with miR-124 NC group,the expression of miR-124,p-PI3K and p-AKT protein in miR-124 mimic group increased,while PTEN mRNA and protein,cell proliferation rate,alkaline phosphatase activity,Col Ⅰ and OPN protein,cell mineralization rate and osteocalcin level decreased,while the pcDNA3.1-PTEN group had the opposite effect(all P<0.05).Compared with miR-124 mimic group,the expressions of p-PI3K and p-AKT protein in miR-124 mimic+pcDNA3.1-PTEN group were decreased,whike PTEN mRNA and protein,cell proliferation rate,alkaline phosphatase activity,Col Ⅰ and OPN protein,cell mineralization rate and osteocalcin level were increased(all P<0.05).Conclusion:MiR-214 regulates the proliferation,differentiation and mineralization of MC3T3-E1 cells by down-regulating PTEN protein expression and activating PI3K/AKT signaling pathway.

参考文献/References:

[1] 史凡凡,赵继荣,马 同,等.中医药治疗糖尿病性骨质疏松症从“脾肾论治”机制[J].陕西中医,2022,43(5):604-607.
[2] 刘 峰,孙庆山,白田雨.温肾强骨方对肿瘤中后期并发骨质疏松症患者骨密度、骨形成及骨吸收的影响[J].陕西中医,2021,42(6):712-715.
[3] Patel D,Wairkar S.Bone regeneration in osteoporosis:Opportunities and challenges[J].Drug Deliv Transl Res,2023,13(2):419-432.
[4] Laird C,Benson H,Williams KA.Pharmacist interventions in osteoporosis management:A systematic review[J].Osteoporos Int,2023,34(2):239-254.
[5] Chakrabarti K,Mccune WJ.Glucocorticoid-induced osteoporosis in premenopausal women:Management for the rheumatologist[J].Curr Opin Rheumatol,2023,35(3):161-169.
[6] Vasikaran SD,Miura M,Pikner R,et al.Practical considerations for the clinical application of bone turnover markers in osteoporosis[J].Calcif Tissue Int,2023,112(2):148-157.
[7] 贺 轲,李 凯.真核基因组编码大量长链非编码RNA癌易感性候选基因15在肺癌细胞中表达上调且通过miR-153-3p激活Wnt/β-catenin通路调控A549细胞增殖、侵袭及化疗敏感性实验研究[J].陕西医学杂志,2022,51(7):776-782.
[8] Zhang K,Liu X,Tang Y,et al.Fluid shear stress promotes osteoblast proliferation and suppresses mitochondrial-mediated osteoblast apoptosis through the miR-214-3p-ATF4 signaling axis[J].Physiol Res,2022,71(4):527-538.
[9] 杨一秋,李 兰,解继胜,等.葛根素介导PTEN-PI3K-AKT信号通路抑制绝经后骨质疏松症的机制研究[J].中国骨质疏松杂志,2022,28(3):347-351.
[10] Cui Y,Li B,Du J,et al.Dibutyl phthalate causes MC3T3-E1 cell damage by increasing ROS to promote the PINK1/Parkin-mediated mitophagy[J].Environ Toxicol,2022,37(10):2341-2353.
[11] Yu S,Luan J,Liu Y,et al.MiR-296 promotes osteoblast differentiation by upregulating cbfal[J].Pharmacology,2020,105(3-4):190-201.
[12] 匡嘉兵,申文娟,马永刚,等.miR-455-3p靶向HIPK2调控高糖环境下成骨细胞的增殖和凋亡[J].中国组织工程研究,2020,24(11):1641-1646.
[13] Liu J,Li D,Dang L,et al.Osteoclastic miR-214 targets TRAF3 to contribute to osteolytic bone metastasis of breast cancer[J].Sci Rep,2017,10(7):40487.
[14] 赵 娜,邱国萍,王满庭.miR-214靶向PTEN介导PI3K/AKT信号通路对糖尿病肾病大鼠的保护机制[J].西部医学,2022,34(3):360-366,374.
[15] 韩纯杰,刘金霞,马慧勇.miR-214-3p通过靶向PTEN抑制骨肉瘤U2OS细胞凋亡的研究[J].长治医学院学报,2017,31(6):411-413.
[16] Sun Y,Zhang H,Meng J,et al.S-palmitoylation of PCSK9 induces sorafenib resistance in liver cancer by activating the PI3K/AKT pathway[J].Cell Rep,2022,40(7):111194.
[17] Hashemi M,Etemad S,Rezaei S,et al.Progress in targeting PTEN/PI3K/Akt axis in glioblastoma therapy:Revisiting molecular interactions[J].Biomed Pharmacother,2023,158:114204.
[18] Yin R,Jiang J,Deng H,et al.MiR-140-3p aggregates osteoporosis by targeting PTEN and activating PTEN/PI3K/AKT signaling pathway[J].Hum Cell,2020,33(3):569-581.
[19] Liu XF,Hou Y,Fu Y,et al.The role of PTEN/PI3K/AKT signaling pathway in apoptosis of liver cells in cocks with manganese toxicity[J].Biol Trace Elem Res,2022,200(10):4444-4452.
[20] Khan KH,Wong M,Rihawi K,et al.Hyperglycemia and phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)inhibitors in phase Ⅰ trials:Incidence,predictive factors,and management[J].Oncologist,2016,21(7):855-860.
[21] 王建武,李晓妮,赵 强,等.调控miR-152对骨质疏松大鼠模型骨生物力学、骨重建平衡及骨组织Hedgehog信号通路的影响[J].陕西医学杂志,2022,51(8):944-947.
[22] 林 丽,张 卓,哈斯达来,等.炎症微环境下miR-30a调控Runx相关转录因子2对人牙周膜干细胞成骨分化的影响[J].陕西医学杂志,2021,50(5):519-524.

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备注/Memo

备注/Memo:
基金项目:内蒙古自治区自然科学基金资助项目(2022MS08012,2022MS08058); 内蒙古自治区卫生健康科技计划项目(202202160); 内蒙古自治区高等学校科学技术研究项目(NJZZ22614); 内蒙古医科大学联合项目(YKD2021LH005,YKD2021LH026); 内蒙古医科大学科技创新团队项目(YKD2022TD034); 内蒙古医科大学附属医院博士科研启动金资助项目(2022NYFYBS003)
更新日期/Last Update: 2023-12-05