外胚层发育不良A1蛋白对类成釉上皮细胞增殖及细胞周期影响的研究  

Effect of ectodysplasin-A1 on proliferation and cell cycle of ameloblast-like cell

作　　者：刘博宇 孔宣婷 路艮其 张国忠 贾娴娴[3] 杜情情 郑书深 郭长军[1] 沈文静[1] Liu Boyu;Kong Xuanting;Lu Genqi;Zhang Guozhong;Jia Xianxian;Du Qingqing;Zheng Shushen;Guo Changjun;Shen Wenjing(Department of Prosthodontics,School and Hospital of Stomatology,Hebei Medical University&Hebei Key Laboratory of Stomatology&Hebei Clinical Research Center for Oral Diseases,Shijiazhuang 050017,China;Department of Forensic Pathology,College of Forensic Medicine,Hebei Medical University,Shijiazhuang 050017,China;Department of Pathogen Biology,Institute of Basic Medicine,Hebei Medical University,Shijiazhuang 050017,China;Department of Stomatology,Xingtai Medical College,Xingtai 054000,China)

机构地区：[1]河北医科大学口腔医学院·口腔医院修复科河北省口腔医学重点实验室河北省口腔疾病临床医学研究中心,石家庄050017 [2]河北医科大学法医学院法医病理教研室,石家庄050017 [3]河北医科大学基础医学院病原生物教研室,石家庄050017 [4]邢台医学高等专科学校口腔系,054000

出　　处：《中华口腔医学杂志》2021年第4期349-354,共6页Chinese Journal of Stomatology

基　　金：河北省科学技术厅科技计划(17277734D);河北省政府资助临床医学优秀人才培养项目(2019061441-2);河北医科大学口腔医院自主培育计划(kq201701)。

摘　　要：目的探讨外胚层发育不良A1(ectodysplasin-A1,EDA1)蛋白对类成釉上皮细胞(ameloblast-like epithelial cell,LS8细胞)增殖和细胞周期的影响。方法用野生型EDA1基因质粒pCR3-Flag-EDA1-W(野生组)、综合征型突变EDA1基因质粒pCR3-Flag-EDA1-H252L(突变组)及空载体质粒pCR3-Flag(对照组)转染LS8细胞,培养0、24、48、72、96 h时噻唑蓝法检测各组细胞增殖活性(A值),流式细胞术检测各组细胞周期,上述实验重复3次。结果培养72 h,与对照组(0.105±0.032)相比,野生组细胞增殖活性(0.201±0.009)显著增加(P<0.05);培养96 h,与对照组(0.168±0.054)及突变组(0.194±0.059)相比,野生组细胞增殖活性(0.386±0.066)显著增加(P<0.05);各时间点突变组与对照组细胞增殖活性差异均无统计学意义(P>0.05)。与对照组(65.4%±2.1%)及突变组(66.6%±3.1%)相比,野生组G0/G1期细胞分布比例(51.2%±1.1%)显著降低(P<0.01);与对照组(23.1%±2.0%)及突变型组(21.9%±1.8%)相比,野生型组S期细胞分布比例(37.3%±2.4%)显著升高(P<0.01);突变组细胞周期细胞分布比例与对照组差异无统计学意义(P>0.05)。结论野生型EDA1基因可促进LS8细胞增殖,并促进G0/G1期细胞向S期转化;综合征型突变EDA1基因(EDA1-H252L)使EDA1蛋白促进细胞增殖和调控细胞周期的功能丧失。Objective To investigate the effects of ectodysplasin-A1(EDA1)on the proliferation and cell cycle of ameloblast-like epithelial cells(LS8 cells).Methods Wild EDA1 plasmid pCR3-Flag-EDA1-W(wild group),syndrome mutant EDA1 plasmid pCR3-Flag-EDA1-H252L(mutant group)and empty vector plasmid pCR3-Flag(control group)were transfected into LS8 cells.Cell proliferation was detected by methyl thiazolyl tetrazolium(MTT)assay and cell cycle was detected by flow cytometry.All tests were repeated three times.Results Compared with the control group(0.105±0.032),the proliferation activity of the wild group(0.201±0.009)was significantly higher after 72 h(P<0.05).Compared with the control group(0.168±0.054)and the mutant group(0.194±0.059),the proliferation activity of the wild group(0.386±0.066)was significantly higher after 96 h(P<0.05).There was no significant difference between the mutant group and the control group at all time points(P>0.05).In the G0/G1 phase,compared with the control group(65.4%±2.1%)and the mutant group(66.6%±3.1%),the cell distribution ratio of the wild group(51.2%±1.1%)was significantly lower(P<0.01).In the S phase,compared with the control group(23.1%±2.0%)and the mutant group(21.9%±1.8%),the cell distribution ratio of the wild type group(37.3%±2.4%)was significantly higher(P<0.01).There was no significant difference in cell cycle distribution between the mutant group and the control group(P<0.05).Conclusions Wild EDA1 promotes the proliferation of LS8 cells and the transformation from G0/G1 to S phase.The syndrome mutant EDA1(EDA1-H252L)loses its function of regulating the cell proliferation and cell cycle of LS8 cells.

关 键 词：外胚叶发育不全蛋白质类 成釉质细胞 上皮细胞 细胞增殖 细胞周期 先天缺牙 

分 类 号：R781[医药卫生—口腔医学]