Linc-POU3F3 is overexpressed in in-stent restenosis patients and induces VSMC phenotypic transformation via POU3F3/miR-449a/KLF4 signaling pathway.

BACKGROUND

With the extensive application of stent implantation in patients undergoing percutaneous coronary interventions (PCI), there are chances that in-stent restenosis (ISR)-a major vascular complication caused by vascular smooth muscle cell (VSMC) phenotypic transformation-might occur.

OBJECTIVES

This study sought to evaluate the role of lincRNA-POU3F3 on VSMC phenotypic transformation and the underlying mechanism.

METHODS

VSMCs were used in our research. We first constructed a gene delivery system through an assembly of lipofectamine and a functional plasmid DNA (pDNA) encoding lincRNA-POU3F3 or MicroRNA-449a, and then, transfected it to VSMCs. Moreover, lentivirus-mediated KLF4 inhibitor (KLF4 siRNA) was also used in these cells. Expression of relevant proteins, such as smooth muscle myosin heavy chain (SM-MHC), alpha smooth muscle actin (α-SMA), osteopontin (OPN), and kruppel-like factor 4 (KLF4), was examined by western blot or immunofluorescence (IF) assay. CCK-8 and wound healing assays were performed to assess the growth and migration of VSMCs. qRT-PCR was used to assess linc-POU3F3 and miR-449a levels. Luciferase reporter assay was also performed.

RESULTS

POU3F3 levels were significantly higher in ISR patients compared to controls. We observed that linc-POU3F3 promoted VSMC proliferation and migration, and induced VSMC phenotypic transformation via POU3F3/miR-449a/KLF4 signaling pathway.

CONCLUSION

Linc-POU3F3 promotes phenotypic transformation of VSMCs via POU3F3/miR-449a/KLF4 pathway. It may provide a theoretical basis to attenuate ISR via pharmacological inhibition of this biomarker or at least serve as a predictor of diagnosis or prognosis of patients with restenosis.