Actin stabilization induces apoptosis in cultured porcine epithelial cell rests of Malassez

作者：Aida N; Ushikubo T; Kobayashi F; Sako R; Suehara M; Furusawa M; Muramatsu T^*

来源：International Endodontic Journal, 2016, 49(7): 663-669.

DOI：10.1111/iej.12494

• actin stabilization
• apoptosis
• epithelial cell rests of Malassez
• jasplakinolide

AimTo test whether actin stabilization by jasplakinolide induces inhibition of cell viability and apoptosis in epithelial cell rests of Malassez (ERM).
MethodologyERM derived from porcine were spread in a 96-well dish (5x10(4)/well) using Dulbecco's modified Eagle's medium. The actin-specific stabilization reagent, jasplakinolide, was incorporated into the culture medium and incubated for 24h. To evaluate cell viability, the WST-1 assay was carried out and absorption (450nm) was measured. To detect apoptotic cells, monoclonal antibody to single-strand DNA (ssDNA) was used and absorption (405nm) was measured. Actin stabilization and apoptosis induced by jasplakinolide were morphologically investigated by staining with Alexa Fluor 568 phalloidin and observed under a fluorescent microscope. As a negative control, DMSO was used instead of jasplakinolide. Differences between the jasplakinolide-treated group and the control group were analysed statistically using the Student's t-test.
ResultsCell viability decreased in a concentration-dependent manner, and cell viability in the jasplakinolide-treated ERM was lower than that in nontreated ERM (n=16, P<0.01). Apoptotic cells in the jasplakinolide-treated ERM were more frequently detected compared to that in nontreated ERM (n=16, P<0.01). Morphologically, shrinkage, irregular forms and fragmentation of nuclei suggesting apoptotic bodies were observed in jasplakinolide-treated ERM, whilst actin filaments were extended in non-treated ERM.
ConclusionActin stabilization by jasplakinolide inhibited cell viability and induced apoptosis in epithelial cell rests of Malassez.

• 出版日期2016-7