ARHGAP29 Regulates Proliferation of Mouse Embryonic Palatal Mesenchymal Cells

doi:10.13701/j.cnki.kqyxyj.2022.03.016

Abstract

Abstract: Objective: To investigate the effect of ARHGAP29 on proliferation of cells from mouse embryonic palatal mesenchymal (EPM). Methods: Immunohistochemical technique was used to confirm the expression of ARHGAP29 in mice palate. The mice EPM cells of GD13.5-day were cultured in vitro. Immunofluorescence technique was used to identify the cellular characteristics. The primary cultured EPM cells were transfected with three pair of Arhgap29-siRNA to establish a gene knockdown cell model. The efficiency of gene knockdown was detected by real-time PCR and western blotting methods. The effects of Arhgap29 gene silencing on the proliferation of EMP cells were assessed by CCK-8 assay. Results: The results of immunohistochemistry showed that ARHGAP29 was significantly expressed in the palate of GD13.5 mice. Immunofluorescence detection showed the strong positive green fluorescence staining of Vimentin in cytoplasm and the negative staining of Cytokeratinc. Real-time PCR and western blotting results indicated that the mRNA and protein level of ARHGAP29 were significantly down-regulated in Arhgap29-siRNA group comparing with blank and negative control, and Arhgap29-siRNA3 showed the best silencing effect (P<0.001). The results of CCK8 assay showed that the proliferation ability of EPM cells began to increase after 48 hours Arhgap29-siRNA3 treatment, and the OD value was significantly increased after 72 hours (P<0.05). Conclusion: Our results indicated that Arhgap29 can promote the proliferation of cells from mouse EMP, which may increase the adhesion of palatal shelves and oral epithelium during the embryonic stage, and affect the palate normal development.

Key words: ARHGAP29, palatal mesenchymal cell, RNA interference, cell proliferation

RUAN Wenyan, HOU Yuzhuan, CHI Dandan, YU Lili, DUAN Xiaohong, HUANG Yongqing. ARHGAP29 Regulates Proliferation of Mouse Embryonic Palatal Mesenchymal Cells[J]. Journal of Oral Science Research, 2022, 38(3): 275-279.

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