Inhibition of Rho-associated kinases disturbs the collective cell migration of stratified TE-10 cells.
BACKGROUND
The collective cell migration of stratified epithelial cells is considered to be an important phenomenon in wound healing, development, and cancer invasion; however, little is known about the mechanisms involved. Furthermore, whereas Rho family proteins, including RhoA, play important roles in cell migration, the exact role of Rho-associated coiled coil-containing protein kinases (ROCKs) in cell migration is controversial and might be cell-type dependent. Here, we report the development of a novel modified scratch assay that was used to observe the collective cell migration of stratified TE-10 cells derived from a human esophageal cancer specimen.


RESULTS
Desmosomes were found between the TE-10 cells and microvilli of the surface of the cell sheet. The leading edge of cells in the cell sheet formed a simple layer and moved forward regularly; these rows were followed by the stratified epithelium. ROCK inhibitors and ROCK small interfering RNAs (siRNAs) disturbed not only the collective migration of the leading edge of this cell sheet, but also the stratified layer in the rear. In contrast, RhoA siRNA treatment resulted in more rapid migration of the leading rows and disturbed movement of the stratified portion.


CONCLUSIONS
The data presented in this study suggest that ROCKs play an important role in mediating the collective migration of TE-10 cell sheets. In addition, differences between the effects of siRNAs targeting either RhoA or ROCKs suggested that distinct mechanisms regulate the collective cell migration in the simple epithelium of the wound edge versus the stratified layer of the epithelium.
Department of Histology and Cell Biology, Yokohama City University School of Medicine, Yokohama, Kanagawa-ken, Japan. zeong3@mac.com.|Department of Plastic and Reconstructive Surgery, Fujisawa Shounandai Hospital, Fujisawa, Kanagawa-ken, Japan. zeong3@mac.com.|Department of Plastic and Reconstructive Surgery, Yokohama City University Hospital, Yokohama, Kanagawa-ken, Japan. zeong3@mac.com.
Taro
T
Mikami
Department of Histology and Cell Biology, Yokohama City University School of Medicine, Yokohama, Kanagawa-ken, Japan. k6652@yokohama-cu.ac.jp.
Keiichiro
K
Yoshida
Department of Histology and Cell Biology, Yokohama City University School of Medicine, Yokohama, Kanagawa-ken, Japan. hajisawada@gmail.com.
Hajime
H
Sawada
Department of Histology and Cell Biology, Yokohama City University School of Medicine, Yokohama, Kanagawa-ken, Japan. eski_mm@yahoo.co.jp.
Michiyo
M
Esaki
Department of Plastic and Reconstructive Surgery, Yokohama City University Hospital, Yokohama, Kanagawa-ken, Japan. burngdy@hotmail.com.
Kazunori
K
Yasumura
Department of Histology and Cell Biology, Yokohama City University School of Medicine, Yokohama, Kanagawa-ken, Japan. micono@mac.com.
Michio
M
Ono
D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D004938:Esophageal Neoplasms; D006801:Humans; D035683:MicroRNAs / Q000502:physiology; D034741:RNA, Small Interfering / Q000494:pharmacology*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*
D035683:MicroRNAs; D034741:RNA, Small Interfering; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases