富组蛋白1对人表皮细胞株HaCaT增殖和迁移功能的影响  

Influence of histatin 1 on the proliferation and migration of HaCaT cells

作　　者：蒋艳[1] 王仙园[1] 罗向东[2] 

机构地区：[1]第三军医大学护理学院,重庆400038 [2]第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室

出　　处：《中华烧伤杂志》2012年第3期207-212,共6页Chinese Journal of Burns

基　　金：国家自然科学基金（30730093）

摘　　要：目的了解富组蛋白1（Hstl）对人表皮细胞株HaCaT增殖、迁移功能的影响方法（1）常规培养HaCaT细胞，按照随机数字表法（分组方法下同）分为对照组与100、30、3g／mLHsll纰以及10ng／ml。重组人表皮生长因子（rhEGF）组、30μg／mLHstl＋10ng／mLrhEGF组，每组样小数为27。对照组不添加刺激因素,后5组分别加入相应浓度Hstl和（或）rhEGF，培养24、48、72h采用细胞计数法检测各组细胞增殖水平。（2）将HaCarr细胞分为对照组与100、30、3μg／mLHstl组。每组样本数为27。对照组不添加刺激因素，后3组分别加入相应浓度Hstl，培养24、48、72h采用流式细胞术检测各组细胞周期，计算增殖指数（PI）。（3）将HaCaT细胞分为对照组、30μg／mLHstl组、10ng／mLrhEGF组、30μg／mLHstl＋10ng／mLrhEGF组、15μg／mLHstl＋5ng／mLrhEGF组、15μg／mLHstl＋10ng／mLrhEGF组，每组样小数为10。对照组不添加刺激因素，后5组分别加入相应浓度的Hstl和（或）rhEGF培养。将各组细胞分为2份，一份用丝裂霭素C处理2h，另一份不作处理,均行划痕实验，划痕后0（即刻）、16、24h观测细胞迁移情况并计t算划痕愈合面积百分比。对数据行方差分析、LSD-t检验或Dunnettt检验。结果（1）培养24h，10ng／mLrhEGF组、30μg／mLHstl＋10ng／mLrhEGF组细胞数显若高于对照组（t值分别为3．813、5．410，P〈0．05或P〈0，01）。与培养48h时30、3μg／mLHstl组比较除外,其余各Hstl和（或）rhEGF对照组培养48、72h细胞数均显著高于对照组（t值为7．754-24．979，P值均小于0．01）。培养72h，100μg／mEHstl组细胞数为（19，21±0．59）×10^4个，明显高于30μg／mLHstl组的（16．19±0．53）×10^4个及3μg／mLHstl组的（15．38±0．13）×10^4个（t值分别为11．391、19．017，P值均小于0．01）；30μg／mLHstl＋10ng／mLrhEGF组细胞数高于30、3μg／mLHstl组及10ng／mlJrhEGF组（t值为4�Objective To study the influence of histatin 1 （ Hstl ） on the proliferation and migra- tion of human epidermal cell line HaCaT. Methods （ 1 ） HaCaT cells were routinely cultured and divided into control group, 100, 30, and 3μg/mL Hstl groups, 10 ng/mL recombinant human epidermal growth factor （rhEGF） group, and 30 μg/mL Hstl ＋ 10 ng/mL rhEGF group, according to the random number ta- ble （ the same dividing method used for following grouping） , with 27 samples in each group. NO stimulating factor was added in control group, while Hstl and （or） rhEGF in corresponding concentration （s） was （were） added in the latter 5 groups. Cell proliferation was assayed by cell counting method at post culture hour （PCH） 24, 48, and 72. （2） HaCaT cells were divided into control group and 100, 30, and 3 μg/mL Hstl groups, with 27 samples in each group. NO stimulating factor was added in control group, while Hstl in corresponding concentration was added in the latter 3 groups. Cell cycle was assayed with flow cytometry at PCH 24, 48, and 72, and P1 was calculated. （3） HaCaT cells were divided into control group, 30 μg/mL Hstl group, 10 ng/mL rhEGF group, 30 μg/mL Hstl ＋ 10 ng/mL rhEGF group, 15 μg/mL Hstl ＋5 ng/mL rhEGF group, and 15μg/mL Hstl ＋10 ng/mL rhEGF group, with 10 samples in each group. NO stimulating factor was added in control group, while Hstl and （or） rhEGF in corresponding con- centration （s） was （were） added in the latter 5 groups. Cells in each group were divided into two portions： cells in one portion were treated by mitomycin C for 2 hours, while ceils in the other portion were not. Scratching assay was conducted in both portions of cells. Cell migration was measured at post scratching hour （PSH） O, 16, and 24, and the wound-area healing rate was calculated. Data were processed with analysis of variance, and LSD- t test or Dunnett t test was applied in paired comparison among groups. Results （ 1 ） At PCH 24, the cell numbers in 10 ng/mL

关 键 词：富组蛋白质类 细胞增殖 细胞运动 创伤和损伤 HACAT细胞 

分 类 号：R96[医药卫生—药理学]