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钙对成釉细胞系ALC细胞生物学特性的影响
引用本文:高震,侯瑞凯,宋索成,阮建平. 钙对成釉细胞系ALC细胞生物学特性的影响[J]. 口腔医学, 2023, 43(1): 39-45. DOI: 10.13591/j.cnki.kqyx.2023.01.007
作者姓名:高震  侯瑞凯  宋索成  阮建平
作者单位:1 西安交通大学口腔医院陕西省颅颌面精准医学研究重点实验室,陕西西安(710004)2 西安交通大学口腔医院预防保健中心第一门诊部,陕西西安(710004)3 西安交通大学金属强度国家重点实验室,陕西西安(710049)4 国家增材制造创新中心,陕西西安(710117)5 陕西省牙颌面疾病临床研究中心,西安交通大学口腔医院口腔预防保健科,陕西西安(710004)
基金项目:国家自然科学基金面上项目(81470034)
摘    要:目的 观察钙对成釉细胞系ALC细胞增殖、凋亡及细胞周期等生物学特性的影响。方法 培养ALC细胞系,在高糖DMEM培养基中配制0、2.0、2.5、3.0、3.5 mmol/L CaCl2溶液,利用倒置显微镜检测在两种培养时间(24、48 h)下ALC细胞形态是否改变。通过CCK-8法分析钙离子对ALC细胞增殖的作用,运用Hoechst染色观察钙离子对ALC细胞凋亡的影响;通过PI染色FCM法分析钙离子对ALC细胞生长周期的干预。利用Western blot检测钙离子对ALC细胞中周期蛋白Cyclin A、Cyclin B、Cyclin D表达的调节。结果 0 mmol/L CaCl2组,ALC细胞形态呈卵圆或多边形,细胞间连接紧密呈铺路石样生长。其他不同浓度的钙干预24、48 h后,ALC细胞形态未见明显变化;CCK-8法结果显示钙干预24、48 h后,随着钙浓度增加,ALC细胞的存活率呈微弱降低趋势,但此趋势无统计学意义;Hoechst染色结果显示不同浓度的钙干预24、48 h后,ALC细胞的凋亡数目随时间延长可见增加,但不同钙离子浓度组间差...

关 键 词:ALC细胞    增殖  凋亡

Effect of calcium on biological properties of the ameloblast ALC
GAO Zhen,HOU Ruikai,SONG Suocheng,RUAN Jianping. Effect of calcium on biological properties of the ameloblast ALC[J]. Stomatology, 2023, 43(1): 39-45. DOI: 10.13591/j.cnki.kqyx.2023.01.007
Authors:GAO Zhen  HOU Ruikai  SONG Suocheng  RUAN Jianping
Affiliation:Key laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi'an Jiaotong University, Xi'an 710004, China
Abstract:Objective To observe the effect of calcium on biological characteristics (proliferation, apoptosis and cell cycle) of ALC ameloblasts. Methods ALC cell lines were cultured in vitro in DMEM medium with high glucose at different concentrations (0, 2.0, 2.5, 3.0 and 3.5 mmol/L CaCl2 aqueous solution) for 24 h and 48 h, respectively. Changes of ALC cells under two kinds of incubation time were observed with an inverted microscope. CCK-8 method was used to analyze the effect of calcium ion on ALC cell proliferation. Hoechst staining was used to observe the effect of calcium ion on ALC cell apoptosis. PI staining and FCM method were used to analyze the effect of calcium ions on the growth cycle of ALC cells. Western blot was used to detect the effect of calcium ions on the expression of Cyclin A, Cyclin B and Cyclin D in ALC cells. Results In the 0 mmol/L CaCl2 group, ALC cells were oval or polygonal in shape, and the cells were closely connected and grew like paving stones. In other concentration groups, the morphology of ALC cells did not change significantly after calcium intervention for 24 h and 48 h. Results of CCK-8 method showed that the survival rate of ALC cells slightly decreased with increasing calcium ions concentration after calcium intervention for 24 h and 48 h. However, there was no significant differences in this trend. Results of Hoechst staining showed that the number of ALC cell apoptosis did not increase significantly after different concentrations of calcium intervention for 24 h and 48 h. With the increase of calcium ion concentration, results of PI staining and FCM method showed that the cell cycle of ALC cells gradually increased in S phase and decreased in G1 and G2 phase gradually. Western blot results showed that the expression of Cyclin A and Cyclin B in ALC cells decreased and the expression of Cyclin D increased after different concentrations of calcium intervention for 24 h and 48 h. Conclusion In this study, calcium has no significant effect on the proliferation and apoptosis of ALC cells. Calcium, however, has an effect on the ALC cell cycle. Results of this study show that calcium ions has no obvious toxic or side effects on the ameloblasts, which could be used to explore the possible mechanism and effect of calcium on dental fluorosis.
Keywords:ALC cells  calcium  proliferation  apoptosis  
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