Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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MicroRNA-200a/210 Controls Proliferation and Osteogenic Differentiation of Human Adipose Tissue Stromal Cells

MicroRNA-200a/210의 인체 지방 유래 중간엽 줄기세포 골분화 및 증식 조절 기전

  • Kim, Young Suk (Department of Physiology, School of Medicine, Pusan National University) ;
  • Park, Hee Jeong (Department of Physiology, School of Medicine, Pusan National University) ;
  • Shin, Keun Koo (Department of Physiology, School of Medicine, Pusan National University) ;
  • Lee, Sun Young (Department of Physiology, School of Medicine, Pusan National University) ;
  • Bae, Yong Chan (Department of Plastic Surgery, School of Medicine, Pusan National University) ;
  • Jung, Jin Sup (Department of Physiology, School of Medicine, Pusan National University)
  • 김영숙 (부산대학교 의학전문대학원 생리학 교실) ;
  • 박희정 (부산대학교 의학전문대학원 생리학 교실) ;
  • 신근구 (부산대학교 의학전문대학원 생리학 교실) ;
  • 이선영 (부산대학교 의학전문대학원 생리학 교실) ;
  • 배용찬 (부산대학교 의학전문대학원 성형외과학 교실) ;
  • 정진섭 (부산대학교 의학전문대학원 생리학 교실)
  • Received : 2017.05.02
  • Accepted : 2017.07.18
  • Published : 2017.07.30
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Abstract

MicroRNAs control the differentiation and proliferation of human adipose tissue-derived stromal cells (hADSCs). However, the role of miR-200a and miR210 on the osteogenic differentiaton of hADSCs has not been determined. hADSCs were isolated from human adipose tissues. Direct binding of mircoRNA to target mRNAs was determined by luciferase assay of the constructs containing putative microRNA binding sites within 3' untranslated region of target mRNAs. Overexpression of miR-200a increased the proliferation and osteogenic differentiation of hADSCs, while causing downregulation of the levels of ZEB2. Inhibition of miR-200a with antisense RNAs inhibited the proliferation and osteogenic differentiation of hADSCs. Overexpression of miR-210 was found to inhibit the proliferation of hADSCs but increase the osteogenic differentiation, while causing downregulation of the levels of IGFBP3. Inhibition of miR-210 with antisense RNAs increased the proliferation but inhibited the osteogenic differentiation of hADSCs. Analysis of the luciferase activity of the constructs containing the miR-200a target site within the ZEB2 3' region and the miR-210 target site within the IGFBP3 3' region revealed lower activity in the miR-200a- or miR-210-transfected hADSCs than in control miRNA-transfected hADSCs. Downregulation of ZEB2 or IGFBP3 in the hADSCs showed similar effects on both their proliferation and osteogenic differentiation with that of miR-200a and miR-210 overexpression, respectively. The results of the current study indicate that miR-200a and miR-210 regulate the osteogenic differentiation and proliferation of hADSCs through the direct targeting of IGFBP3 and ZEB2, respectively.

MicroRNA는 인체 지방 유래 중간엽 줄기세포(hADSC)의 분화와 증식을 조절할 수 있으나 hADSC에서 miR-200a와 miR210의 역할은 아직까지 보고된 바가 없다. 표적 mRNA의 3' UTR 에 결합할 수 있는 construct를 이용한 luciferase assay를 통하여 microRNA가 직접적으로 표적 mRNA에 결합하는 것을 확인 하였다. hADSC에서 miR-200a 과발현은 ZEB2의 발현 감소를 통해 hADSC의 분화와 증식을 증가시키는 것을 확인할 수 있었으며, miR210의 과발현은 IGFBP3의 발현 감소를 통해 hADSC의 증식을 감소시키는 반면, 분화는 증가시키는 것을 확인 할 수 있었다. hADSC에서 miR210의 발현 억제는 표적유전자의 발현을 증가시킴으로써 hADSC의 증식을 증가시키는 반면, 분화를 억제시키는 것을 확인할 수 있었다. luciferase assay통하여 microRNA-200a/210의 과발현이 대조군에 비해 표적 유전자인 ZEB2/ IGFBP3의 luciferase 활성화를 감소시키는 것을 확인하였으며, hADSC에서 ZEB2/ IGFBP3 발현 억제는 microRNA-200a/210 과발현과 유사한 효과를 나타내는 것을 확인할 수 있었다. 이러한 결과는 microRNA-200a/210가 hADSC의 분화와 증식을 각각의 표적 유전자인 ZEB2/ IGFBP3을 통해 조절한다는 것을 나타낸다.

Keywords

References

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