Effect of calcium ion regulating KLK4 expression on the growth of ameloblast
10.12016/j.issn.2096-1456.2024.00.00
- VernacularTitle:钙离子调控KLK4表达对成釉细胞生长的影响
- Author:
Xiaojing LIU
1
;
Meili GAO
;
Jianping RUAN
Author Information
1. 榆林市第一医院口腔科,陕西 榆林(719000)
- Keywords:
ameloblast;
ALC cells;
calcium ion;
kallikrein-4;
cell growth;
cell viability;
cell cycle;
cell apoptosis;
glucose-regulated protein 78
- From:
Journal of Prevention and Treatment for Stomatological Diseases
2024;32(10):746-755
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of calcium ions on the expression of kallikrein-4(KLK4)and cell growth of ameloblast,and to provide an experimental basis for calcium ion promoting normal mineralization of enamel.Methods ALC cells were treated with 0,2.0,2.5,3.0,and 3.5 mmol/L CaCl2 for 24 and 48 h.KLK4 expression was analyzed by qRT-PCR and Western blot analysis.The viability of ALC cells was determined by using CCK-8.Annex-inV-FITC/PI dual staining combined with flow cytometry and Hoechst 33342 staining were used to detect the ALC cell cycle and cell apoptosis.The protein expression level of glucose-regulated protein 78(GRP78)was measured by West-ern blot analysis.Results After 24 h of treatment with 2.5,3.0,and 3.5 mmol/L CaCl2,the expression of KLK4 mRNA was increased(P＜0.05),and after 24 h of treatment with 2.0,2.5,3.0,and 3.5 mmol/L CaCl2,the expression of KLK4 protein was increased(P＜0.05).After 48 h of treatment with 3.0 mmol/L and 3.5 mmol/L CaCl2,the expression of KLK4 mRNA and protein was increased(P＜0.05).Compared with the control group,the viability of ALC cells was in-creased after 24 and 48 h of treatment with 2.0,2.5,and 3.0 mmol/L CaCl2(P＜0.05),and the highest cell viability was observed with 2.5 mmol/L CaCl2.Hoechst 33342 staining results showed that 3.0 mmol/L and 3.5 mmol/L CaCl2 may promote apoptosis in ALC cells.Flow cytometry showed that the proportion of G2/M phase cells and the apoptosis rate increased after 3.5 mmol/L CaCl2 treatment for 24 h(P＜0.05),compared with the 0,2.0,2.5,and 3.0 mmol/L CaCl2 groups.After 24 h of treatment with 3.0 mmol/L and 3.5 mmol/L CaCl2,the expression of GRP78 protein was re-duced(P＜0.05),and after 48 h of treatment with 2.5 mmol/L CaCl2,the expression of GRP78 protein was reduced(P＜0.05).Conclusion Calcium ions can promote the increase of KLK4 expression and cell viability in ALC cells,but a higher concentration of calcium ions can block the G2/M phase of ALC cells,thus inducing apoptosis of ALC cells and reducing the expression of apoptosation-related protein GRP78.
- Full text:2024101414152493337钙离子调控KLK4表达对成釉细胞生长的影响.pdf
