title: RNA Interference-based Investigation of the Function of Heat Shock Protein 27 during Corneal Epithelial Wound Healing.
abstract: Small interfering RNA (siRNA) is among the most widely used RNA interference methods for the short-term silencing of protein-coding genes. siRNA is a synthetic RNA duplex created to specifically target a mRNA transcript to induce its degradation and it has been used to identify novel pathways in various cellular processes. Few reports exist regarding the role of phosphorylated heat shock protein 27 (HSP27) in corneal epithelial wound healing. Herein, cultured human corneal epithelial cells were divided into a scrambled control-siRNA transfected group and a HSP27-specific siRNA-transfected group. Scratch-induced directional wounding assays, and western blotting, and flow cytometry were then performed. We conclude that HSP27 has roles in corneal epithelial wound healing that may involve epithelial cell apoptosis and migration. Here, step-by-step descriptions of sample preparation and the study protocol are provided.
authors: Department of Ophthalmology, Saevit Eye Hospital.
authors: Aeri
authors: A
authors: Yoo
authors: Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center.
authors: Hyun-Min
authors: HM
authors: Park
authors: Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center.
authors: Soon-Suk
authors: SS
authors: Kang
authors: Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center.
authors: Eun-Soon
authors: ES
authors: Kim
authors: Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center.
authors: Hungwon
authors: H
authors: Tchah
authors: Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center; jykim2311@amc.seoul.kr.
authors: Jae Yong
authors: JY
authors: Kim
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques / Q000379:methods; D055551:HSP27 Heat-Shock Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006360:Heat-Shock Proteins; D006801:Humans; D018832:Molecular Chaperones; D010766:Phosphorylation; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D014162:Transfection / Q000379:methods; D014945:Wound Healing / Q000502:physiology*
chemical_list: D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D012333:RNA, Messenger; D034741:RNA, Small Interfering