Integrin β1 mediates gingipain - induced cell cycle arrest in osteoblasts

doi:10.3877/cma.j.issn.1674-1366.2018.04.003

Abstract

Abstract:

Objective

To investigate the effect of integrin β1 (Itgb1) in mediating gingipain-induced cell cycle arrest in osteoblasts.

Methods

MC3T3-E1 cells were treated with 8.348 U/L gingipains from 12 to 72 h. Cell counting kit-8 (CCK-8) assay and Propidium Iodide/Rnase (PI/Rnase) staining were used to detect the proliferation activity and cell cycle distribution of G₀/G₁, S, G₂/M phases. The expression of cyclin D1 and CDK4 were evaluated by western blotting after gingipain treatment for 12 h. MC3T3-E1 cells were transducted with lentivirus encoding mouse Itgb1 to overexpress Itgb1. All data were analyzed by SPSS 22.0 software package.

Results

CCK-8 assay showed that 8.348 U/L gingipain treatment inhibited cellular proliferation from 12 to 72 h. The percentage of cells in G₀/G₁ phase was increased from (62.0 ± 2.0) % to (88.2 ± 2.3) % (F = 35.218, P<0.001) , and the percentage of cells in S phase was decreased from (36.8 ± 6.2) % to (5.9 ± 2.3) % (F= 33.980, P<0.001) compared to control cells. Western blotting showed the protein expression of cyclin D1 and CDK4 were reduced by about 65.0% (F_{cyclin D1}= 60.294, P_{cyclin D1}<0.001) and 40.3% (F_CDK4= 19.212, P_CDK4= 0.002) compared to control cells. Meanwhile, gingipains reduced Itgb1 protein level, overexpression of Itgb1 partially reversed gingipain-induced cell cycle arrest. The percentage of cells in S phase was significantly higher in LV-Itgb1 cells (14.4 ± 3.1) % than that in LV-Ctrl cells (5.8 ± 1.1) % upon gingipain treatment (F= 39.226, P= 0.017) . Similar effects of overexpression of Itgb1 were observed in cyclin D1 and CDK4 down-regulation (F_{cyclin D1}= 61.740, P_{cyclin D1}= 0.033; F_CDK4= 41.635, P_CDK4= 0.014) .

Conclusion

Itgb1 is involved in mediating cell cycle arrest induced by gingipains in osteoblasts.

Key words: Integrin beta1, Cell cycle, Osteoblasts, Chronic periodontitis, Gingipains, Porphyromonas gingivalis

Na Xu, Juan Wu, Qihong Qiu, Fuping Zhang, Min Liang. Integrin β1 mediates gingipain - induced cell cycle arrest in osteoblasts[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2018, 12(04): 213-220.

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References 35

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