中国医科大学学报

中国医科大学学报
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中国医科大学学报 ›› 2018, Vol. 47 ›› Issue (11): 1015-1020.doi: 10.12007/j.issn.0258-4646.2018.11.013

• 论著 • 上一篇    下一篇

OTX2基因表达下调对MC3T3-E1细胞增殖分化和自噬的影响

田玉楼, 于默, 史娅婕, 张薇   

  1. 中国医科大学口腔医学院正畸教研室, 沈阳 110002
  • 收稿日期:2018-05-09 出版日期:2018-11-30 发布日期:2018-12-06
  • 通讯作者: 田玉楼 E-mail:yltian@cmu.edu.cn
  • 作者简介:田玉楼(1969-),女,教授,博士.
  • 基金资助:
    沈阳市科学技术计划(F15-199-1-55)

Effect of OTX2 Downregulation on Proliferation and Autophagy in MC3T3-E1 Cells

TIAN Yulou, YU Mo, SHI Yajie, ZHANG Wei   

  1. Department of Orthodontics, School of Stomatology, China Medical University, Shenyang 110002, China
  • Received:2018-05-09 Online:2018-11-30 Published:2018-12-06

摘要: 目的 研究OTX2基因对小鼠成骨细胞系MC3T3-E1增殖分化及自噬水平的影响。方法 应用OTX2基因小干扰RNA(siRNA)转染MC3T3-E1细胞,采用MTT法测定细胞增殖情况,流式细胞仪检测细胞周期及凋亡水平,化学比色法检测细胞碱性磷酸酶(ALP)合成情况,单丹磺酰尸胺(MDC)荧光染色观察各组自噬体表达情况。结果 与对照组比较,实验组细胞增殖抑制率明显升高(P<0.05),S期细胞百分比减少,S期细胞比例以及细胞的增殖指数显著降低(P<0.05),凋亡率无统计学差异(P>0.05);ALP活性明显低于对照组(P<0.05);实验组可见少量MDC点状荧光,荧光细胞比例降低,阴性对照组点状荧光较多,实验组与阴性对照组荧光细胞比例有统计学差异(P<0.05)。结论 OTX2基因表达下调可以抑制MC3T3-E1细胞增殖,降低细胞分化水平,引起MC3T3-E1细胞自噬水平降低。

关键词: OTX2, siRNA, 转染, 自噬

Abstract: Objective To study the effects of the downregulation of OTX2 on proliferation,differentiation,and autophagy in MC3T3-E1 cells. Methods Small interfering RNA (siRNA) targeting the OTX2 gene was transfected into MC3T3-E1 cells. Cell proliferation was determined by the MTT assay. Flow cytometry was used to detect the cell cycle phase and apoptosis. The activity of alkaline phosphatase (ALP) was determined by a colorimetric assay. Cell autophagy was observed by monodansylcadaverine (MDC) staining. Results The inhibition rate of the experimental group was significantly higher than the control group (P<0.05). The percentage of cells in S phase in the experimental group decreased after transfection,and the ratio of cells in S phase and the cell proliferation index were less than that of the control group (P<0.05);the cell apoptosis results showed that there was no significant difference between the experimental and the control groups (P>0.05). The ALP activity in the experimental group was lower than that observed in the control group (P<0.05). MDC fluorescence staining showed that the staining intensity in the experimental group was weak with little point structure,while the control group showed higher staining intensity. Conclusion The downregulation of the OTX2 gene can inhibit proliferation and reduce the levels of differentiation and autophagy in MC3T3-E1 cells.

Key words: OTX2 gene, siRNA, cell transfection, autophagy

中图分类号: 

  • R329.28
[1] 陈彦泽,史册. 调控下颌骨发育的转录因子家族的研究进展[J]. 国际口腔医学杂志,2012,39(6):751-755. DOI:10.3969/j.issn. 1673-5749.2012.06.014.
[2] BALLESTA-MARTÍNEZ MJ,LÓPEZ-GONZÁLEZ V,DULCET LA,et al. Autosomal dominant oculoauriculovertebral spectrum and 14q23.1 microduplication[J]. Am J Med Genet A,2013,161(8):2030-2035. DOI:10.1002/ajmg.a.36007.
[3] SONG C,TONG F. Autophagy induction is a survival response against oxidative stress in bone marrow-derived mesenchymal stromal cells[J]. Cytotherapy,2014,16(10):563-565. DOI:10.1016/j.jcyt.2014.04.006.
[4] CARAMES B,HASEGAWA A,TANIGUCHI N,et a1. Autophagy activation by rapamycin reduces severity of experimental osteoarthritiss[J]. Ann Rheum Dis,2012,71(4):575-581. DOI:10.1136/annrheumdis-2011-200557.
[5] WU J,NIU J,LI X,et al. Hypoxia induces autophagy of bone marrow-derived mesenchymal stem cells via activation of ERK1/2. Cell Physiol Biochem,2014,33(5):1467-1474. DOI:10.1159/000358711.
[6] DONG W,GE J,ZHANG P,et al. Phenotypic characterization of craniofacial bone marrow stromalcells:unique properties of enhanced osteogenesis,cell recruitment,autophagy,and apoptosis resistance[J]. Cell Tissue Res,2014,358(1):165-175. DOI:10.1007/s00441-014-1927-4.
[7] MENG HZ,ZHANG WL,LIU F,et al. Advanced glycation end products affect osteoblast proliferation and function by modulating autophagy via the receptor of advanced glycation end products/raf protein/mitogen-activated protein kinase/extracellular signal-regulated kinase kinase/extracellular signal-regulated kinase (RAGE/Raf/MEK/ERK) pathway[J]. J Biol Chem,2015,290(47):28189-28199. DOI:10.1074/jbc.M115.669499.
[8] WEI M,DUAN D,LIU Y,et al. Autophagy may protect MC3T3-E1 cell from fluoride-induced apoptosis[J]. Mol Med Rep,2014,9(6):2309-2315. DOI:10.3892/mmr.2014.2079.
[9] LÜ XH,ZHAO DH,CAI SZ,et al. Autophagy plays a protective role in cell death of osteoblasts exposure to lead chloride[J]. Toxicol Lett,2015,239(2):131-140. DOI:10.1016/j.toxlet.2015.09.014.
[10] 王雅雯. 自噬基因Beclin1在肿瘤中的作用及其与口腔癌的关系[J]. 中国实用口腔科杂志,2011,4(6):374-376. DOI:1674-1595(2011) 06-0374-03.
[11] GREENHILL C. Bone:autophagy regulates bone growth in mice[J]. Nat Rev Endocrinol,2016,12(1):4. DOI:10.1038/nrendo.2015.214.
[12] SHAPIRO IM,LAYFIELD R,LOTZ M,et al. Boning up on autophagy:the role of autophagy in skeletal biology[J]. Autophagy,2014,10(1):7-19. DOI:10.4161/auto.26679.
[13] MAHEU ME,RESSLER KJ. Developmental pathway genes and neural plasticity underlying emotional learning and stress-related disorders[J]. Learn Mem,2017,24(9):492-501. DOI:10.1101/lm.044271.116.
[14] TAKENOUCHI T,NISHINA S,KOSAKI R,et al. Concurrent deletion of BMP4 and OTX2 genes,two master genes in ophthalmogenesis[J]. Eur J Med Genet,2013,56(1):50-53. DOI:10.1016/j.ejmg.2012.10.007.
[15] SU Z,ZHANG Y,LIAO B,et al. Antagonism between the transcription factors NANOG and OTX2 specifies rostral or caudal cell fate during neural patterning transition[J]. J Biol Chem,2018,293(12):4445-4455. DOI:10.1074/jbc.M117.815449.
[16] ZHENG M,JIAO L,TANG X,et al. Tau haploin sufficiency causes prenatal loss of dopaminergic neurons in the ventral tegmental area and reduction of transcription factor orthodenticle homeobox 2 expression[J]. FASEB J,2017,31(8):3349-3358. DOI:10.1096/fj.201601303R.
[17] LEE HHC,BERNARD C,YE Z,et al. Genetic Otx2 mis-localization delays critical period plasticity across brain regions[J]. Mol Psychiatry,2017,22(5):680-688. DOI:10.1038/mp.2017.1.
[18] 谢丽敏,刘丹丹,宗菲,等. 不同浓度的抗菌肽RISE-AP12对体外培养成骨细胞生物活性的影响[J]. 口腔医学,2017,37(2):114-120. DOI:10.13591/j.cnki.kqyx.2017.02.005.
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