摘要
目的观察微小RNA-26b(mi R-26b)对不同分化状态神经干细胞(NSCs)系C17. 2向肝细胞生长因子(HGF)趋化迁移的影响。方法 (1)HGF刺激下不同分化状态NSCs细胞mi R-26b检测取C17. 2细胞,置于含N2和F12的诱导分化培养基中诱导分化。将细胞分为A、B、C、D组,A、B、C组分别于诱导分化12、24、72 h三个时间点终止诱导分化,D组为诱导分化前的细胞。采用实时荧光定量PCR法检测各组细胞mi R-26b。将诱导分化培养基更换为含25 ng/m L的HGF完全培养基,孵育5 h,每组均设相应对照,予无HGF的完全培养基孵育5 h。检测各组细胞mi R-26b。(2)抑制、过表达mi R-26b对C17. 2细胞向HGF的趋化迁移的影响取对数生长期C17. 2细胞,分为两组,分别转染mi R-26b mimic(促进mi R-26b表达)、mi R-26b NC(对照),将完全生长培养基更换为含N2和F12的诱导分化培养基,分别于诱导分化0、12、24、72 h时终止诱导分化,采用Transwell实验测算各分化时间细胞的迁移细胞率。取对数生长期C17. 2细胞,分为两组,分别转染mi R-26b inhibitor(抑制mi R-26b表达)、mi R-26b乱序对照(ConⅠ),将完全生长培养基更换为含N2和F12的诱导分化培养基,分别于诱导分化0、12、24、72 h时终止诱导分化,采用Transwell实验测算各分化时间细胞的迁移细胞率。结果 A、B、C、D组细胞mi R-26b相对表达量分别为1. 16±0. 07、1. 39±0. 11、2. 30±0. 11、1. 00±0. 03,与D组比较,B、C组胞mi R-26b相对表达量升高,P均<0. 05。A、B、C、D组HGF处理5 h时细胞mi R-26b相对表达量分别为2. 02±0. 43、2. 28±0. 35、2. 50±0. 41、2. 21±0. 23,A、B、C、D组未经处理细胞mi R-26b相对表达量分别为1. 21±0. 12、1. 47±0. 21、2. 30±0. 20、1. 00±0. 04,与未经处理者比较,HGF处理5 h时A、B、D组细胞mi R-26b相对表达量均升高,P均<0. 05。与转染NC者比较,分化0、12、24、72 h时转染mi R-26b mimic的细胞迁移率均升高(P均<0. 05)。与转染ConⅠ者比较,�
Objective To observe the effects of miR-26b on chemotactic migration of C17.2 neural stem cells of different differentiation stats toward hepatocyte growth factor(HGF).Methods The expression of miR-26b in neural stem cells(NSCs)of different differentiated states was detected after stimulation with HGF.C17.2 cells were placed on medium containing N2 and F12 for induced differentiation.The cells were divided into groups A,B,C and D.The induced differentiation was terminated in the groups A,B and C at 12,24 and 72 h,respectively,while the cells in the group D was undifferentiated cells.The miR-26b of each group was detected by real-time quantitative PCR.The induced differentiation medium was replaced with a complete medium containing 25 ng/mL HGF and incubated for 5 h,while each group was given a corresponding control group and incubated with complete medium without HGF for 5 h.The expression of miR-26b in each group was detected.C17.2 cells in the logarithmic phrase were divided into two groups and were transfected with miR-26b mimics(which promoted miR-26b expression)and miR-26b NC(controls),respectively.After that,the complete growth medium was replaced by the induction differentiation medium containing N2 and F12.Cells were cultured on the induction medium for 0,12,24 and 72 h,respectively.Transwell assay was used to detect the migration rate of C17.2 cells of different differentiation states.We used the same methods above,and inhibited the expression of miR-26b of neural stem cells through transfecting miR-26b inhibitor(which inhibited miR-26b expression)and con I(controls),and then we measured the migration rate of neural stem cells of different differentiation toward HGF.Results The relative expression levels of miR-26b in the groups A,B,C and D were 1.16±0.07,1.39±0.11,2.30±0.11,and 1.00±0.03,respectively.Compared with the group D,the relative expression level of miR-26b in cells of groups B and C increased,and significant difference was found between them(P<0.05).After 5-hour HGF treatment,the relative
作者
熊英
汪显耀
秦臻
田羽
王岚
潘际刚
许键炜
XIONG Ying;WANG Xianyao;QIN Zhen;TIAN Yu;WANG Lan;PAN Jigang;XU Jianwei(Basic Medical College of Guizhou Medical University,Guiyang 550025,China)
出处
《山东医药》
CAS
2018年第43期1-5,共5页
Shandong Medical Journal
基金
贵州省科技厅科技计划项目(黔科合LH字2016-7361)
贵州省科技厅联合基金项目(黔科合LH字-2015-7325)
