口腔疾病防治2024，Vol.32Issue(10)：746-755,10.DOI:10.12016/j.issn.2096-1456.2024.00.00
钙离子调控KLK4表达对成釉细胞生长的影响
Effect of calcium ion regulating KLK4 expression on the growth of ameloblast
摘要
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.关键词
成釉细胞/ALC细胞/钙离子/激肽释放酶4/细胞生长/细胞活力/细胞周期/细胞凋亡/葡萄糖调节蛋白78Key words
ameloblast/ALC cells/calcium ion/kallikrein-4/cell growth/cell viability/cell cycle/cell apoptosis/glucose-regulated protein 78分类
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刘晓静,高美丽,阮建平..钙离子调控KLK4表达对成釉细胞生长的影响[J].口腔疾病防治,2024,32(10):746-755,10.基金项目
陕西省重点研发计划项目(2023-YBSF-659) （2023-YBSF-659）
榆林市科技计划项目(YF-2021-44) This study was supported by the grants from Shaanxi Province Key Research and Development Program(No.2023-YBSF-659) （YF-2021-44）
Yulin Science and Technology Project(No.YF-2021-44). （No.YF-2021-44）
