摘要
目的探究敲低microRNA(miR).19a和miR-19b表达对人脑胶质瘤细胞系SNB19细胞生物学特征的影响。方法常规培养SNB19细胞,脂质体介导miR-19a、miR-19b抑制物转染SNB19细胞,同时设未转染组(对照组1、无义序列转染组和miR-19a+miR-19b抑制物转染组。应用RT.PCR检测转染后细胞miR-19a、miR-19b的表达,MTT、流式细胞术分别检测转染后细胞增殖能力和细胞周期的变化,Transwell实验检测转染后细胞的侵袭能力。结果与对照组和无义序列转染组比较,抑制物转染组细胞miR-19a和miR-19b的表达、增殖和侵袭能力均降低,差异有统计学意义(P〈O.05),而且miR-19a+miR-19b抑制物转染组细胞miR.19a和miR-19b的表达、转染后2、3、4、5d细胞的增殖能力、侵袭能力均低于miR-19a、miR-19b抑制物转染组.差异有统计学意义(JD〈0.05);抑制物转染组较对照组和无义序列转染组细胞周期延迟,且miR-19a+miR-19b抑制物转染组较miR-19a、miR-19b抑制物单独转染组细胞延迟更为明显。结论miR-19a和miR-19b可能是癌微RNA.有望作为人脑胶质瘤基因治疗的侯选靶点。
objective To investigate the effects of knocking down ofmiR-19a and miR-19b on the biological characteristics of SNB19 glioblastoma cells. Methods Oligonucleotides inhibitor of miR-19a and miR-19b (miR-19a inhibitor or miR-19b inhibitor) mediated by lipofectamine2000 were transfected to SNB19 cells to knock down miR-19a and miR-19b; control group (without transfection), group D (performing transfection with nonsense sequence) and group E (performing transfection with both miR-19a inhibitor and miR-19b inhibitor) were established. Real time PCR was conducted to detect the expressions ofmiR-19a and miR-19b in these groups after the transfection. The cell proliferation rate and cell cycle kinetics were detected by 3-(4, 5-Dime- -thylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively; the cell invasive ability was evaluated by Transwell assay. Results As compared with those in control group and group D, the expressions of miR-19a and miR-19b, proliferation activity and invasive ability of cells in the miR-19a/19b inhibitor transfected cells (group A/B) were significantly reduced (P〈0.05). The expressions of miR-19a and miR-19b and the proliferation activity and invasive ability of cells 2, 3, 4 and 5 d after the transfection in group E were significantly reduced as compared with those in group A/B (P〈0.05). Delayed cell cycle in group A/B and group E was noted as compared with that in control group and group D; and group E enjoyed more obviously delayed cell cycle than group A/B (P〈0.05). Conclusion MiR-19a and miR-19b might be oncomiRs, and may be candidate target miRNAs for gene therapy of glioma.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2011年第4期365-368,共4页
Chinese Journal of Neuromedicine