shRNA knockdown of integrin-linked kinase on hPDLCs migration, proliferation, and apoptosis under cyclic tensile stress

Abstract

Objective: To investigate the roles of integrin-linked kinase (ILK) in mediating the cell migration, proliferation, and apoptosis of human periodontal ligament cells (hPDLCs) in response to cyclic tensile stress.

Methods: Primary hPDLCs were obtained through the enzyme digestion and tissue culture method. Short hairpin ILK-expressing hPDLCs were constructed using a recombinant lentiviral vector that specifically targeted ILK gene expression. The silencing of the ILK gene was identified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The hPDLCs were seeded on a flexible substrate and loaded with cyclic tensile stress at 0.5 Hz for 0, 2, 4, and 8 hr, consecutively, with the Flexcell Tension System. The response of cell migration was tested by the scratch assay. Cell proliferation was characterized by optical density (OD) value of cell counting kit-8 (CCK-8) test and Ki67 mRNA expression of qRT-PCR. Cell apoptosis was determined by flow cytometry and Caspase-3 mRNA expression of qRT-PCR.

Results: Knocking down ILK substantially reduces migration and proliferation as well as regulates the sensitivity of hPDLCs to apoptosis under cyclic tensile stress.

Conclusions: ILK can promote the proliferation and migration as well as inhibit apoptosis of hPDLCs under cyclic tensile stress.

Keywords: cyclic tensile stress; human periodontal ligament cells; integrin-linked kinase; mechanical signal transduction.