过表达KLF4对小鼠成骨细胞成骨分化的影响

杨佳佳; 刘苹; 徐倩; 刘志华; 李晓明; 汪超; 张波; 邓蔓菁

doi:10.16016/j.1000-5404.201610196

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过表达KLF4对小鼠成骨细胞成骨分化的影响

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- 过表达KLF4对小鼠成骨细胞成骨分化的影响
- Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro
- 第三军医大学学报 2017年39卷第7期页码：628-634
- 作者机构：
- 作者简介：
- 基金信息：
  
  创伤、烧伤与复合伤国家重点实验室自主研究课题(SKLZZ201507)
- DOI：10.16016/j.1000-5404.201610196
  中图分类号：
- 收稿：2016-10-27，
  
  纸质出版：2017-07
- 稿件说明：
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杨佳佳, 刘苹, 徐倩, 等. 过表达KLF4对小鼠成骨细胞成骨分化的影响[J]. 第三军医大学学报, 2017,39(7):628-634.

Jiajia Yang, Ping Liu, Qian Xu, et al. Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro[J]. Journal of Third Military Medical University, 2017, 39(7): 628-634.
杨佳佳, 刘苹, 徐倩, 等. 过表达KLF4对小鼠成骨细胞成骨分化的影响[J]. 第三军医大学学报, 2017,39(7):628-634. DOI： 10.16016/j.1000-5404.201610196.

Jiajia Yang, Ping Liu, Qian Xu, et al. Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro[J]. Journal of Third Military Medical University, 2017, 39(7): 628-634. DOI： 10.16016/j.1000-5404.201610196.

摘要

Abstract

Objective

To determine the effect of recombinant adenovirus-mediated Krüppel like factor 4 (KLF4) transfection on the osteogenic differentiation of osteoblasts

in vitro

Methods

Methods Primary mouse osteoblasts were isolated from newborn C57 mice by enzymatic digestion and then cultured and identified. Alizarin Red S staining was carried out after osteogenic induction for 21 d. The recombinant adenoviral vector carrying the KLF4 gene (Ad-KLF4) was constructed by pHBAd-EF1-MCS-GFP system. The transfection efficiency was detected by measuring the different multiplicity of infection (MOI) with flow cytometry. Osteoblasts were randomly assigned into control group (non-transfection)

GFP group (Ad-GFP transfection) and KLF4 group (Ad-KLF4 transfection). The expression of KLF4 protein was detected by Western blotting. MTT assay was used to analyze cell proliferation. Real-time PCR was used to detect the expression of osteogenic genes alkaline phosphatase (ALP)

bone sialoprotein (BSP) and runt-related transcription factor2 (RUNX2).

Results

Primary osteoblasts were triangle and polygon in shape under inverted phase contrast micr

oscope. A large number of red calcified nodules were found after Alizarin Red S staining. Restriction enzyme digestion and gene sequencing confirmed Ad-KLF4 was constructed successfully. At the optimal multiplicity of infection (MOI

100 PFU/mL)

Ad-KLF4 transfection resulted in significant increases in protein and mRNA levels of KLF4 (

0.01)

obviously increase in osteoblasts proliferation (

0.05

0.01)

and decreases in the mRNA levels of osteogenic genes ALP

BSP and RUNX2 (

0.05

0.01)

when compared with the control group and GFP group.

Conclusion

Adenovirus-mediated KLF4 over-expression can promote osteoblast proliferation and inhibit osteoblast osteogenic differentiation

and thus inhibit bone formation.

关键词

Keywords

references

Khosla S Oursler M J Monroe D G Estrogen and the skeleton Trends Endocrinol Metab 2012 23 11 576 581 10.1016/j.tem.2012.03.008 Khosla S, Oursler M J, Monroe D G. Estrogen and the skeleton[J]. Trends Endocrinol Metab, 2012, 23(11): 576-581. DOI:10.1016/j.tem.2012.03.008

Sobacchi C Schulz A Coxon F P Osteopetrosis: genetics, treatment and new insights into osteoclast function Nat Rev Endocrinol 2013 9 9 522 536 10.1038/nrendo.2013.137 Sobacchi C, Schulz A, Coxon F P, et al. Osteopetrosis: genetics, treatment and new insights into osteoclast function[J]. Nat Rev Endocrinol, 2013, 9(9): 522-536. DOI: 10.1038/nrendo.2013.137

Vestergaard P Rejnmark L Mosekilde L Selective serotonin reuptake inhibitors and other antidepressants and risk of fracture Calcif Tissue Int 2008 82 2 92 101 10.1007/s00223-007-9099-9 Vestergaard P, Rejnmark L, Mosekilde L. Selective serotonin reuptake inhibitors and other antidepressants and risk of fracture[J]. Calcif Tissue Int, 2008, 82(2): 92-101. DOI:10.1007/s00223-007-9099-9

Gothard D Smith E L Kanczler J M In Vivo Assessment of Bone Regeneration in Alginate/Bone ECM Hydrogels with Incorporated Skeletal Stem Cells and Single Growth Factors PLoS ONE 2015 10 12 e0145080 10.1371/journal.pone.0145080 Gothard D, Smith E L, Kanczler J M, et al. In Vivo Assessment of Bone Regeneration in Alginate/Bone ECM Hydrogels with Incorporated Skeletal Stem Cells and Single Growth Factors[J ] . PLoS ONE, 2015, 10(12): e0145080. DOI:10.1371/journal.pone.0145080

Bouvard B Abed E Yéléhé-Okouma M Hypoxia and vitamin D differently contribute to leptin and dickkopf-related protein 2 production in human osteoarthritic subchondral bone osteoblasts Arthritis Res Ther 2014 16 5 459 10.1186/s13075-014-0459-3 Bouvard B, Abed E, Yéléhé-Okouma M, et al. Hypoxia and vitamin D differently contribute to leptin and dickkopf-related protein 2 production in human osteoarthritic subchondral bone osteoblasts[J]. Arthritis Res Ther, 2014, 16(5): 459. DOI:10.1186/s13075-014-0459-3

McConnell B B Yang V W Mammalian Krüppel-like factors in health and diseases Physiol Rev 2010 90 4 1337 1381 10.1152/physrev.00058.2009 McConnell B B, Yang V W. Mammalian Krüppel-like factors in health and diseases[J]. Physiol Rev, 2010, 90(4): 1337-1381. DOI:10.1152/physrev.00058.2009

Evans P M Liu C Roles of Krüpel-like factor 4 in normal homeostasis, cancer and stem cells Acta Biochim Biophys Sin (Shanghai) 2008 40 7 554 564 10.1111/j.1745-7270.2008.00439.x Evans P M, Liu C. Roles of Krüpel-like factor 4 in normal homeostasis, cancer and stem cells[J]. Acta Biochim Biophys Sin (Shanghai), 2008, 40(7): 554-564. DOI:10.1111/j.1745-7270.2008.00439.x

Sung M T Hsu H T Lee C C Krüppel-like factor 4 modulates the migration and invasion of hepatoma cells by suppressing TIMP-1 and TIMP-2 Oncol Rep 2015 34 1 439 446 10.3892/or.2015.3964 Sung M T, Hsu H T, Lee C C, et al. Krüppel-like factor 4 modulates the migration and invasion of hepatoma cells by suppressing TIMP-1 and TIMP-2[J]. Oncol Rep, 2015, 34(1): 439-446. DOI:10.3892/or.2015.3964

Kim J H Kim K Youn B U Krüppel-like factor 4 attenuates osteoblast formation, function, and cross talk with osteoclasts J Cell Biol 2014 204 6 1063 1074 10.1083/jcb.201308102 Kim J H, Kim K, Youn B U, et al. Krüppel-like factor 4 attenuates osteoblast formation, function, and cross talk with osteoclasts[J]. J Cell Biol, 2014, 204(6): 1063-1074. DOI:10.1083/jcb.201308102

Song C Z Keller K Murata K Functional interaction between coactivators CBP/p300, PCAF, and transcription factor FKLF2 J Biol Chem 2002 277 9 7029 7036 10.1074/jbc.M108826200 Song C Z, Keller K, Murata K, et al. Functional interaction between coactivators CBP/p300, PCAF, and transcription factor FKLF2[J]. J Biol Chem, 2002, 277(9): 7029-7036. DOI:10.1074/jbc.M108826200

Tien Y T Chang M H Chu P Y Downregulation of the KLF4 transcription factor inhibits the proliferation and migration of canine mammary tumor cells Vet J 2015 205 2 244 253 10.1016/j.tvjl.2014.12.031 Tien Y T, Chang M H, Chu P Y, et al. Downregulation of the KLF4 transcription factor inhibits the proliferation and migration of canine mammary tumor cells[J]. Vet J, 2015, 205(2): 244-253. DOI:10.1016/j.tvjl.2014.12.031

Yoshida T Yamashita M Hayashi M Krüppel-like factor 4 contributes to high phosphate-induced phenotypic switching of vascular smooth muscle cells into osteogenic cells J Biol Chem 2012 287 31 25706 25714 10.1074/jbc.M112.361360 Yoshida T, Yamashita M, Hayashi M. Krüppel-like factor 4 contributes to high phosphate-induced phenotypic switching of vascular smooth muscle cells into osteogenic cells[J]. J Biol Chem, 2012, 287(31): 25706-25714. DOI:10.1074/jbc.M112.361360

Moore D L Blackmore M G Hu Y KLF family members regulate intrinsic axon regeneration ability Science 2009 326 5950 298 301 10.1126/science.1175737 Moore D L, Blackmore M G, Hu Y, et al. KLF family members regulate intrinsic axon regeneration ability[J]. Science, 2009, 326(5950): 298-301. DOI:10.1126/science.1175737

Yoshida T Gan Q Owens G K Krüppel-like factor 4, Elk-1, and histone deacetylases cooperatively suppress smooth muscle cell differentiation markers in response to oxidized phospholipids Am J Physiol Cell Physiol 2008 295 5 C1175 C1182 10.1152/ajpcell.00288.2008 Yoshida T, Gan Q, Owens G K. Krüppel-like factor 4, Elk-1, and histone deacetylases cooperatively suppress smooth muscle cell differentiation markers in response to oxidized phospholipids[J]. Am J Physiol Cell Physiol, 2008, 295(5): C1175-C1182. DOI:10.1152/ajpcell.00288.2008

Michikami L Fukushi T Tanaka M Krüppel-like factor 4 regulates membranous and endochondral ossification Exp Cell Res 2012 318 4 311 325 10.1016/j.yexcr.2011.12.013 Michikami L, Fukushi T, Tanaka M, et al. Krüppel-like factor 4 regulates membranous and endochondral ossification[J]. Exp Cell Res, 2012, 318(4): 311-325. DOI: 10.1016/j.yexcr.2011.12.013

Komori T Regulation of osteoblast differentiation by Runx2 Adv Exp Med Biol 2010 658 43 49 10.1007/978-1-4419-1050-9_5 Komori T. Regulation of osteoblast differentiation by Runx2[J]. Adv Exp Med Biol, 2010, 658: 43-49. DOI:10.1007/978-1-4419-1050-9_5

Marinovich R Soenjaya Y Wallace G Q The role of bone sialoprotein in the tendon-bone insertion Matrix Biol 2016 52 54 10.1016/j.matbio.2016.01.016 Marinovich R, Soenjaya Y, Wallace G Q, et al. The role of bone sialoprotein in the tendon-bone insertion[J]. Matrix Biol, 2016, 52-54: 325-338. DOI:10.1016/j.matbio.2016.01.016

Fakhry M Hamade E Badran B Molecular mechanisms of mesenchymal stem cell differentiation towards osteoblasts World J Stem Cells 2013 5 4 136 148 10.4252/wjsc.v5.i4.136 Fakhry M, Hamade E, Badran B, et al. Molecular mechanisms of mesenchymal stem cell differentiation towards osteoblasts[J]. World J Stem Cells, 2013, 5(4):136-148. DOI: 10.4252/wjsc.v5.i4.136

Takahashi K Yamanaka S Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors Cell 2006 126 4 663 676 10.1016/j.cell.2006.07.024 Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J]. Cell, 2006, 126(4): 663-676. DOI:10.1016/j.cell.2006.07.024

Saulnier N Puglisi M A Lattanzi W Gene profiling of bone marrow-and adipose tissue-derived stromal cells: a key role of Krüppel-like factor 4 in cell fate regulation Cytotherapy 2011 13 3 329 340 10.3109/14653249.2010.515576 Saulnier N, Puglisi M A, Lattanzi W, et al. Gene profiling of bone marrow-and adipose tissue-derived stromal cells: a key role of Krüppel-like factor 4 in cell fate regulation[J]. Cytotherapy, 2011, 13(3): 329-340. DOI:10.3109/14653249.2010.515576

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