Inhibitory effects of ameloblastin on epithelial cell proliferation.

Abstract:

OBJECTIVE:Ameloblastin is an enamel matrix protein expressed in several tissues. Many potential mechanisms have been identified by which ameloblastin functions as an extracellular matrix protein. However, the biological effects of ameloblastin on gingival epithelial cells remain unclear. In the present study, we established a novel system to purify recombinant human ameloblastin and clarified its biological functions in epithelial cells in vitro. DESIGN:Recombinant human ameloblastin was isolated from COS-7 cells overexpressing HaloTag-fused human ameloblastin by the HaloTag system and then purified further by reverse-phase high-performance liquid chromatography. SCC-25 cells, derived from human oral squamous cell carcinoma, were treated with recombinant ameloblastin and then cell survival was assessed by a WST-1 assay. Cell cycle analysis was performed by flow cytometry. RESULTS:The novel purification system allowed effective recovery of the recombinant ameloblastin proteins at a high purity. Recombinant ameloblastin protein was found to suppress the proliferation of SCC-25 cells. Flow cytometric analysis showed that ameloblastin treatment induced cell cycle arrest G1 phase. CONCLUSIONS:We developed a procedure for production of highly purified recombinant human ameloblastin. Biological analyses suggest that ameloblastin induces cell cycle arrest in epithelial cells and regulates the progression of periodontitis.

journal_name

Arch Oral Biol

journal_title

Archives of oral biology

authors

Saito N,Ariyoshi W,Okinaga T,Kamegawa M,Matsukizono M,Akebiyama Y,Kitamura C,Nishihara T

doi

10.1016/j.archoralbio.2014.05.010

subject

Has Abstract

pub_date

2014-08-01 00:00:00

pages

835-40

issue

8

eissn

0003-9969

issn

1879-1506

pii

S0003-9969(14)00112-5

journal_volume

59

pub_type

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