HPF1 regulates tendon stem/progenitor cell senescence and tendon repair via PARP1-mediated poly-ADP ribosylation of HuR

Han Weifeng,GU Dongqiang,Chen Hongguang,Tao Xu,Chen Lei 한국유전학회 2024 Genes & Genomics Vol.46 No.1

Background Tendon stem/progenitor cells (TSPCs) play a vital role in tendon repair, regeneration and homeostasis. However, the specific mechanism of TSPCs aging is still unclear. Objective This study aims to explore the role and molecular mechanism of HPF1 in the aging of TSPCs. Methods Young and aged TSPCs (Y-TSPCs and A-TSPCs) were acquired from 3 to 4 and 24–26-month-old Sprague–Dawley male rats, TSPCs (Y-TSPCs and A-TSPCs) were subjected to senescence-associated β-galactosidase (SA-β-Gal))staining and telomerase activity detection, p16, p21, Scx, Tnmd, Col1, Col3HPF1 and PAPR1 expression levels were detected by Western blot or Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR), Reciprocal co-immunoprecipitation (co-IP) was used to explore the interaction between HPF1 and PARP1. Ribonucleoprotein immunoprecipitation (RNP-IP) was used to analyze the binding of HuR to the senescence marker gene mRNAs, IP was used to perform HPF1 to the PARylation of HuR, and the half-life of p16 and p21 were detected. Finally, we established an in vivo model, and the tendon tissue was used to perform hematoxylin and eosin (HE) and masson’s trichrome staining, as well as the immunohistochemical analysis of Col I and TNMD. Results Compared with Y-TSPCs, A-TSPCs had significantly enhanced cell senescence and significantly reduced tendon differentiation ability, and significantly increased the expression of HPF1 and PARP1. In addition, HPF1 and PARP1 interacted and coordinated the senescence and differentiation of TSPCs, HPF1 could also regulate the expression of p21 and p21, the interaction of p16 or p21 with HuR, and the poly-ADP ribosylation of PARP1 to HuR. HPF1 overexpression and siHuR co-transfection significantly reduced the half-life of p16 and p21, and HPF1 and PARP1 regulated the mRNA levels of p16 and p21 through HuR. Finally, in vivo experiments have shown that HPF1 or PARP1 overexpression could both inhibit the ability of tendon differentiation and promote cell senescence. Conclusions HPF1 promoted the senescence of TSPCs and inhibits the tendon differentiation of TSPCs through PARP1-mediated poly-ADP ribosylation of HuR. Similar content being viewed by others Background Tendon stem/progenitor cells (TSPCs) play a vital role in tendon repair, regeneration and homeostasis. However, the specific mechanism of TSPCs aging is still unclear. Objective This study aims to explore the role and molecular mechanism of HPF1 in the aging of TSPCs. Methods Young and aged TSPCs (Y-TSPCs and A-TSPCs) were acquired from 3 to 4 and 24–26-month-old Sprague–Dawley male rats, TSPCs (Y-TSPCs and A-TSPCs) were subjected to senescence-associated β-galactosidase (SA-β-Gal))staining and telomerase activity detection, p16, p21, Scx, Tnmd, Col1, Col3HPF1 and PAPR1 expression levels were detected by Western blot or Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR), Reciprocal co-immunoprecipitation (co-IP) was used to explore the interaction between HPF1 and PARP1. Ribonucleoprotein immunoprecipitation (RNP-IP) was used to analyze the binding of HuR to the senescence marker gene mRNAs, IP was used to perform HPF1 to the PARylation of HuR, and the half-life of p16 and p21 were detected. Finally, we established an in vivo model, and the tendon tissue was used to perform hematoxylin and eosin (HE) and masson’s trichrome staining, as well as the immunohistochemical analysis of Col I and TNMD. Results Compared with Y-TSPCs, A-TSPCs had significantly enhanced cell senescence and significantly reduced tendon differentiation ability, and significantly increased the expression of HPF1 and PARP1. In addition, HPF1 and PARP1 interacted and coordinated the senescence and differentiation of TSPCs, HPF1 could also regulate the expression of p21 and p21, the interaction of p16 or p21 with HuR, and the poly-ADP ribosylation of PARP1 to HuR. HPF1 overexpression and siHuR co-transfection significantly reduced the half-life of p16 and p21, and HPF1 and PARP1 regulated the mRNA levels of p16 and p21 through HuR. Finally, in vivo experiments have shown that HPF1 or PARP1 overexpression could both inhibit the ability of tendon differentiation and promote cell senescence. Conclusions HPF1 promoted the senescence of TSPCs and inhibits the tendon differentiation of TSPCs through PARP1-mediated poly-ADP ribosylation of HuR. Similar content being viewed by others

A new linear cutting experimental machine for disc-cutter rock breaking with high-pressure water jets assisted

Weifeng Han,Jie Fu,Xingchen Luo,Lu Guo,Yi-Min Xia,Shan He 대한토목학회 2023 KSCE Journal of Civil Engineering Vol.27 No.6

Aiming at the low rock-breaking efficiency of TBM disc cutters in tough rock environments, A new experimental machine combined with the high-pressure water jet rock-breaking method was designed. It can flexibly change the cutting parameters of the water jet, such as spray position, spray height, pressure, etc. Moreover, the three-direction load of the disc cutter during the rock-breaking process can be obtained accurately by strain testing. The rock-breaking experiment reached the following conclusions. The experimental machine can meet the design function. In this study, the normal and rolling forces when the disc cutter breaks rock are significantly lower than the traditional, 26.90% and 28.39%, respectively. The lateral force increased slightly, about 31.17%. Although the energy consumption of water-jet-assisted is higher than the traditional rock-breaking method, the volume of broken rock per unit of time is approximately 5.85 times that of the traditional. It is feasible to increase the energy cost to increase the excavation speed. On the rock surface after the water jet experiment, a rock ridge appears on the rolling trajectory of the disc cutter, which is not conducive to further breaking the rock. The cutting depth of the water jet needs to be adjusted to reduce the height of the rock ridge.

Stability Evaluation of Highway Cut Slope Based on Remote Monitoring of Axial Force of Anchor and Displacement

( Weifeng Sun ),( Changgen Yan ),( Wei Xu ),( Jiangbo Xu ),( Han Bao ),( Yongli Xie ) 대한지질공학회 2019 대한지질공학회 학술발표회논문집 Vol.2019 No.2

During the construction and operation period of highway cut slope, to evaluate the stability of the slope, it is of great necessity to monitor the responses of the slope and its reinforced structures. Compared with the traditional manual monitoring methods of slope, remote monitoring has become the development trend to obtain a timely feedback of potential hazards. This paper presents a case of highway cut slope based on remote monitoring of axial force of anchor and displacement, and the stability of the slope was evaluated. Reinforcement meters and deep displacement sensors were respectively used to monitor the axial forces of anchors and relative displacements of the slope in the In-situ test, and a network cloud platform was used to remotely manage the monitoring data. The results indicate that the axial forces of anchors varies slowly with the increasing of monitoring time within a limited range under the seasonal temperature change, and the displacements tends to be stable except in the winter, indicating the slope is stable and the monitoring is effective. For a highway cut slope with a long length, to remote monitoring the slope with an economical and effective way, it is suggested to install a small number of sensors on the sensitive positions.

Potential Antitumor Activity of SIM-89 in Non-Small Cell Lung Cancer Cells

Jun Pei,Baohui Han,Tianqing Chu,Minhua Shao,Jiajun Teng,Huifang Sha,Aiqing Gu,Rong Li,Jialin Qian,Weifeng Mao,Ying Li 연세대학교의과대학 2017 Yonsei medical journal Vol.58 No.3

Purpose: c-Met and its ligand, hepatocyte growth factor (HGF), play a critical role in oncogenesis and metastatic progression. The aim of this study was to identify inhibited enzymogram and to test the antitumor activity of SIM-89 (a c-Met receptor tyrosine kinaseinhibitor) in non-small cell lung cancer. Materials and Methods: Z’-LYTE kinase assay was employed to screen the kinase enzymogram, and mechanism of action (MOA) analysis was used to identify the inhibited kinases. Cell proliferation was then analyzed by CCK8 assay, and cell migration was determinedby transwell assay. The gene expression and the phosphorylation of c-Met were examined by realtime-PCR and western blotting, respectively. Finally, the secretion of HGF was detected by ELISA assay. Results: c-Met, activated protein kinase (AMPK), and tyrosine kinase A (TRKA) were inhibited by SIM-89 with the IC50 values of 297 nmol/L, 1.31 μmol/L, and 150.2 nmol/L, respectively. SIM-89 exerted adenosine triphosphate (ATP) competitive inhibition on c-Met. Moreover, the expressions of STAT1, JAK1, and c-Met in H460 cells were decreased by SIM-89 treatment, and c-Met phosphorylationwas suppressed in A549, H441, H1299, and B16F10 cells by the treatment. In addition, SIM-89 treatment significantly decreased the level of HGF, which accounted for the activation of c-Met receptor tyrosine kinase. Finally, we showed cell proliferationinhibition and cell migration suppression in H460 and H1299 cells after SIM-89 treatment. Conclusion: In conclusion, SIM-89 inhibits tumor cell proliferation, migration and HGF autocrine, suggesting it’s potential antitumoractivity.