PMID- 36530027
OWN - NLM
STAT- MEDLINE
DCOM- 20230201
LR  - 20230202
IS  - 1899-5276 (Print)
IS  - 1899-5276 (Linking)
VI  - 32
IP  - 1
DP  - 2023 Jan
TI  - CircHIPK3 promotes bone microvascular endothelial cell proliferation, migration 
      and angiogenesis by targeting miR-7 and KLF4/VEGF signaling in steroid-induced 
      osteonecrosis of the femoral head.
PG  - 43-55
LID - 10.17219/acem/153042 [doi]
AB  - BACKGROUND: Circular RNA homeodomain interacting protein kinase 3 (circHIPK3) has 
      been implicated in facilitating angiogenesis in various conditions. However, its 
      role in steroid-induced osteonecrosis of the femoral head (ONFH) remains unclear. 
      OBJECTIVES: To investigate whether circHIPK3 promotes bone microvascular activity 
      and angiogenesis by targeting miR-7 and Kruppel-like factor 4 (KLF4)/vascular 
      endothelial growth factor (VEGF) signaling in ONFH. MATERIAL AND METHODS: Fifty 
      patients with steroid-induced ONFH undergoing hip-preserving surgery or total hip 
      arthroplasty were included in this study. The expression of circHIPK3, miR-7 and 
      KLF4 was evaluated using reverse transcription polymerase chain reaction (RT-PCR) 
      in necrotic and healthy samples of the femoral head. Bone microvascular 
      endothelial cells (BMECs) were extracted and cultured with 0.1 mg/mL 
      hydrocortisone to create a hormonally deficient cell model. These BMECs were then 
      transfected with either circHIPK3 overexpressing or silencing plasmids with or 
      without miR-7 mimics. The MTT assays were used to detect cell proliferation. 
      Scratch assays were used to assess the migration ability of the BMECs. The tube 
      formation was carried out using an in vitro Matrigel angiogenesis assay. Annexin 
      V-FITC/PI and terminal deoxynucleotidyl transferase dUTP nick end labeling 
      (TUNEL) assays were used to assess the degree of apoptosis. Western blot assays 
      were carried out to discern KLF4 and VEGF expression. The interactions of 
      circHIPK3, miR-7 and KLF4 were confirmed using luciferase, RNA-binding protein 
      immunoprecipitation (RIP), RNA pull-down, and fluorescence in situ hybridization 
      (FISH) assays. RESULTS: The circHIPK3 and KLF4 expression was decreased, whereas 
      miR-7 expression was increased in necrotic tissues compared to non-necrotic 
      samples. Both circHIPK3 and KLF4 expression correlated negatively with miR-7. The 
      overexpression of circHIPK3 promoted the proliferative, migratory and angiogenic 
      capabilities of the BMECs, while adding an miR-7 mimic reversed these effects. At 
      the same time, the overexpression of circHIPK3 reduced the apoptosis rate of the 
      BMECs and increased KLF4 and VEGF protein expression, but adding an miR-7 mimic 
      reversed these effects. The FISH, RNA pull-down, RIP, and luciferase assays 
      revealed an interaction between circHIPK3, miR-7 and KLF4. CONCLUSIONS: The 
      circHIPK3 promotes BMEC proliferation, migration and angiogenesis by targeting 
      miR-7 and KLF4/VEGF signaling.
FAU - Peng, Peng
AU  - Peng P
AD  - Department of Orthopedic Surgery, The First Affiliated Hospital of Guangzhou 
      University of Chinese Medicine, China.
AD  - Department of Orthopedic Surgery, Zhuhai Hospital of Integrated Traditional 
      Chinese and Western Medicine, China.
FAU - He, Wei
AU  - He W
AD  - Department of Orthopedic Surgery, The First Affiliated Hospital of Guangzhou 
      University of Chinese Medicine, China.
FAU - Zhang, Yi-Xi
AU  - Zhang YX
AD  - Department of Ophthalmology, Zhuhai Hospital of Integrated Traditional Chinese 
      and Western Medicine, China.
FAU - Liu, Xiao-Hua
AU  - Liu XH
AD  - Department of Orthopedic Surgery, Zhuhai Hospital of Integrated Traditional 
      Chinese and Western Medicine, China.
FAU - Chen, Zhen-Qiu
AU  - Chen ZQ
AD  - Department of Orthopedic Surgery, The First Affiliated Hospital of Guangzhou 
      University of Chinese Medicine, China.
FAU - Mao, Ji-Gang
AU  - Mao JG
AD  - Department of Orthopedic Surgery, Zhuhai Hospital of Integrated Traditional 
      Chinese and Western Medicine, China.
LA  - eng
PT  - Journal Article
PL  - Poland
TA  - Adv Clin Exp Med
JT  - Advances in clinical and experimental medicine : official organ Wroclaw Medical 
      University
JID - 101138582
RN  - 0 (Vascular Endothelial Growth Factor A)
RN  - 0 (Kruppel-Like Factor 4)
RN  - 0 (MicroRNAs)
RN  - 0 (RNA, Circular)
RN  - 0 (Steroids)
RN  - 0 (MIRN7 microRNA, human)
SB  - IM
MH  - Humans
MH  - Vascular Endothelial Growth Factor A/metabolism
MH  - Endothelial Cells
MH  - Kruppel-Like Factor 4
MH  - Femur Head
MH  - In Situ Hybridization, Fluorescence
MH  - *MicroRNAs/metabolism
MH  - Cell Proliferation
MH  - RNA, Circular/genetics
MH  - *Osteonecrosis/metabolism
MH  - Steroids/metabolism
OTO - NOTNLM
OT  - bone microvascular endothelial cell
OT  - circHIPK3
OT  - miR-7
OT  - osteonecrosis of the femoral head
EDAT- 2022/12/20 06:00
MHDA- 2023/02/02 06:00
CRDT- 2022/12/19 02:12
PHST- 2022/12/20 06:00 [pubmed]
PHST- 2023/02/02 06:00 [medline]
PHST- 2022/12/19 02:12 [entrez]
AID - 10.17219/acem/153042 [doi]
PST - ppublish
SO  - Adv Clin Exp Med. 2023 Jan;32(1):43-55. doi: 10.17219/acem/153042.