中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2015年
1期
60-65
,共6页
叶敏华%张安玲%吴雷%王坤%王广秀%贾志凡%黄强%祝新根
葉敏華%張安玲%吳雷%王坤%王廣秀%賈誌凡%黃彊%祝新根
협민화%장안령%오뢰%왕곤%왕엄수%가지범%황강%축신근
微RNAs%RNA,反义%神经胶质瘤%细胞增殖%肿瘤侵润%体外研究
微RNAs%RNA,反義%神經膠質瘤%細胞增殖%腫瘤侵潤%體外研究
미RNAs%RNA,반의%신경효질류%세포증식%종류침윤%체외연구
MicroRNAs%RNA,antisense%Glioma%Cell proliferation%Neoplasm invasiveness%In vitro
目的 探讨抑制miR-106b过表达对人脑胶质瘤细胞株增殖和侵袭的影响.方法 培养人脑U251、LN229及TJ905胶质瘤细胞,并将其分为细胞对照组,转染阴性对照组(简称阴性对照组),转染miR-106b inhibitors组(简称106b inh组).将反义miR-106b用脂质体转染于人脑胶质瘤细胞中,抑制miR-106b的表达.采用实时定量PCR法检测转染后胶质瘤细胞miR-106b的表达,流式细胞术检测细胞周期的变化、噻唑兰比色分析法检测细胞增殖能力、软琼脂克隆形成实验评价细胞非锚定依赖性生长能力及Transwell实验检测细胞侵袭能力的变化.结果 与细胞对照组,阴性对照组比较,(1) 106b inh组3种胶质瘤细胞中miR-106b的表达均被明显抑制;细胞周期进展被抑制,S期细胞比例减少,G0/G1期细胞比例增加,其中U251、TJ905细胞的周期时相分布差异有统计学意义(P值均＜0.05),LN229细胞的周期时相分布差异无统计学意义;(2) 106b inh组转染抑制后48 h,3种胶质瘤细胞的增殖能力均明显减弱,P＜ 0.05;(3)细胞的克隆形成能力也被明显抑制[克隆形成率分别为,U251:(10.6±0.9)％、(10.5±1.2)％对比(3.3±0.4)％;LN229:(12.5±1.7)％、(11.0±2.4)％对比(4.1±0.3)％;TJ905:(9.9±1.2)％、(10.2±0.6)％对比(3.5±0.4)％.P值均＜0.01];(4)穿过Transwell小室的细胞数明显降低[U251:(90.6±5.7)、(85.2±6.1)个对比(27.6±4.0)个;LN229:(22.6±3.7)、(21.2±3.7)个对比(2.4±1.5)个;TJ905:(79.4±4.4)、(76.8±5.9)个对比(11.8±3.2)个.P值均＜0.0l].结论 下调miR-106b的表达可抑制胶质瘤细胞的增殖、生长和侵袭能力.
目的 探討抑製miR-106b過錶達對人腦膠質瘤細胞株增殖和侵襲的影響.方法 培養人腦U251、LN229及TJ905膠質瘤細胞,併將其分為細胞對照組,轉染陰性對照組(簡稱陰性對照組),轉染miR-106b inhibitors組(簡稱106b inh組).將反義miR-106b用脂質體轉染于人腦膠質瘤細胞中,抑製miR-106b的錶達.採用實時定量PCR法檢測轉染後膠質瘤細胞miR-106b的錶達,流式細胞術檢測細胞週期的變化、噻唑蘭比色分析法檢測細胞增殖能力、軟瓊脂剋隆形成實驗評價細胞非錨定依賴性生長能力及Transwell實驗檢測細胞侵襲能力的變化.結果 與細胞對照組,陰性對照組比較,(1) 106b inh組3種膠質瘤細胞中miR-106b的錶達均被明顯抑製;細胞週期進展被抑製,S期細胞比例減少,G0/G1期細胞比例增加,其中U251、TJ905細胞的週期時相分佈差異有統計學意義(P值均＜0.05),LN229細胞的週期時相分佈差異無統計學意義;(2) 106b inh組轉染抑製後48 h,3種膠質瘤細胞的增殖能力均明顯減弱,P＜ 0.05;(3)細胞的剋隆形成能力也被明顯抑製[剋隆形成率分彆為,U251:(10.6±0.9)％、(10.5±1.2)％對比(3.3±0.4)％;LN229:(12.5±1.7)％、(11.0±2.4)％對比(4.1±0.3)％;TJ905:(9.9±1.2)％、(10.2±0.6)％對比(3.5±0.4)％.P值均＜0.01];(4)穿過Transwell小室的細胞數明顯降低[U251:(90.6±5.7)、(85.2±6.1)箇對比(27.6±4.0)箇;LN229:(22.6±3.7)、(21.2±3.7)箇對比(2.4±1.5)箇;TJ905:(79.4±4.4)、(76.8±5.9)箇對比(11.8±3.2)箇.P值均＜0.0l].結論 下調miR-106b的錶達可抑製膠質瘤細胞的增殖、生長和侵襲能力.
목적 탐토억제miR-106b과표체대인뇌효질류세포주증식화침습적영향.방법 배양인뇌U251、LN229급TJ905효질류세포,병장기분위세포대조조,전염음성대조조(간칭음성대조조),전염miR-106b inhibitors조(간칭106b inh조).장반의miR-106b용지질체전염우인뇌효질류세포중,억제miR-106b적표체.채용실시정량PCR법검측전염후효질류세포miR-106b적표체,류식세포술검측세포주기적변화、새서란비색분석법검측세포증식능력、연경지극륭형성실험평개세포비묘정의뢰성생장능력급Transwell실험검측세포침습능력적변화.결과 여세포대조조,음성대조조비교,(1) 106b inh조3충효질류세포중miR-106b적표체균피명현억제;세포주기진전피억제,S기세포비례감소,G0/G1기세포비례증가,기중U251、TJ905세포적주기시상분포차이유통계학의의(P치균＜0.05),LN229세포적주기시상분포차이무통계학의의;(2) 106b inh조전염억제후48 h,3충효질류세포적증식능력균명현감약,P＜ 0.05;(3)세포적극륭형성능력야피명현억제[극륭형성솔분별위,U251:(10.6±0.9)％、(10.5±1.2)％대비(3.3±0.4)％;LN229:(12.5±1.7)％、(11.0±2.4)％대비(4.1±0.3)％;TJ905:(9.9±1.2)％、(10.2±0.6)％대비(3.5±0.4)％.P치균＜0.01];(4)천과Transwell소실적세포수명현강저[U251:(90.6±5.7)、(85.2±6.1)개대비(27.6±4.0)개;LN229:(22.6±3.7)、(21.2±3.7)개대비(2.4±1.5)개;TJ905:(79.4±4.4)、(76.8±5.9)개대비(11.8±3.2)개.P치균＜0.0l].결론 하조miR-106b적표체가억제효질류세포적증식、생장화침습능력.
Objective To study the effect of miR-106b inhibitor on glioma cell line growth and invasive ability in vitro.Methods The miR-106b inhibitor was transfected by liposome to repress the expression of miR-106b,qRT-PCR was taken to measure miR-106b expression after tansfection.The cell cycle kinetics and cell proliferation rate were detected by floweytometry and MTT assay,the cell malignant proliferative ability was evaluated by soft agar assay,and the invasive ability was detected by transwell assay.Results The antisense oligonucleotide of miR-106b effectively down-regulated miR-106b expression in glioma cells.The progression of cell cycle was blocked.The rate of S phase decreased,and the rate of G0/G1 phase increased.The distribution of cell cycle was significantly different in U251 and TJ905 (P ＜ 0.05),but not in LN229.After transfection for 48h,the proliferation activity of three cell lines was obviously inhibited.Also,the cell clone formation was repressed [The rate of clone efficiency:U251:(10.6 ± 0.9)％、(10.5 ± 1.2)％ vs.(3.3 ± 0.4)％;LN229:(12.5 ± 1.7)％、(11.0 ± 2.4)％ vs.(4.1 ± 0.3)％;TJ905:(9.9 ± 1.2)％、(10.2 ± 0.6)％ vs.(3.5 ± 0.4)％.P ＜ 0.01].The invasive ability was also significantly repressed in cells transfected with miR-106b inhibitor as compared to those of the cells transfected with negative control oligonucleotide and control cells [The cell number per view was U251:(90.6±5.7),(85.2±6.1) vs.(27.6±4.0);LN229:(22.6 ±3.7),(21.2±3.7) vs.(2.4 ± 1.5); T J905:(79.4 ± 4.4),(76.8 ± 5.9) vs.(11.8 ± 3.2).P ＜ 0.01].Conclusions Suppression of miR-106b expression could inhibit the proliferative and invasive ability of glioma cells.
