中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2010年
2期
199-203
,共5页
张明芳%郭亚%齐元麟%郑志父
張明芳%郭亞%齊元麟%鄭誌父
장명방%곽아%제원린%정지부
Notch1%RNA干扰%慢病毒%细胞增殖%无胸腺小鼠%种植瘤
Notch1%RNA榦擾%慢病毒%細胞增殖%無胸腺小鼠%種植瘤
Notch1%RNA간우%만병독%세포증식%무흉선소서%충식류
Notch1%RNA interference%lentivirus%cell proliferation%athymic mice%neoplasm transplantation
目的 探讨沉默Notch1基因对人神经胶质瘤U251细胞增殖能力的影响.方法 采用Notch1-shRNA慢病毒感染人胶质瘤U251细胞,筛选稳定低表达Notch1的细胞单克隆,Western blot法检测细胞Notch1蛋白的表达;CCK-8法测定细胞生长;平板克隆形成实验和软琼脂克隆形成实验检测细胞的集落形成能力;裸鼠种植瘤模型观察Notch1沉默对种植瘤生长的影响.结果 成功筛选得到Notch1稳定沉默的U251细胞单克隆,其Notch1蛋白下调(65.3±5.1)%.Notch1稳定沉默细胞的生长增殖能力明显受到抑制,生长曲线与对照组相比明显减慢;Notch1沉默组的平板克隆形成率为(18.6±2.5)%,低于对照组(37.0±3.3)%;Notch1沉默组软琼脂集落形成率(51±5)%,低于对照组(80±4)%;Notch1沉默组裸鼠腋下种植瘤生长速度减慢,30 d后瘤重(0.31±0.09)g,明显低于对照组(2.35±0.71)g;统计学分析差异均有显著性.结论 RNAi沉默Notch1基因可致人胶质瘤U251细胞的体外增殖和体内成瘤能力下降.
目的 探討沉默Notch1基因對人神經膠質瘤U251細胞增殖能力的影響.方法 採用Notch1-shRNA慢病毒感染人膠質瘤U251細胞,篩選穩定低錶達Notch1的細胞單剋隆,Western blot法檢測細胞Notch1蛋白的錶達;CCK-8法測定細胞生長;平闆剋隆形成實驗和軟瓊脂剋隆形成實驗檢測細胞的集落形成能力;裸鼠種植瘤模型觀察Notch1沉默對種植瘤生長的影響.結果 成功篩選得到Notch1穩定沉默的U251細胞單剋隆,其Notch1蛋白下調(65.3±5.1)%.Notch1穩定沉默細胞的生長增殖能力明顯受到抑製,生長麯線與對照組相比明顯減慢;Notch1沉默組的平闆剋隆形成率為(18.6±2.5)%,低于對照組(37.0±3.3)%;Notch1沉默組軟瓊脂集落形成率(51±5)%,低于對照組(80±4)%;Notch1沉默組裸鼠腋下種植瘤生長速度減慢,30 d後瘤重(0.31±0.09)g,明顯低于對照組(2.35±0.71)g;統計學分析差異均有顯著性.結論 RNAi沉默Notch1基因可緻人膠質瘤U251細胞的體外增殖和體內成瘤能力下降.
목적 탐토침묵Notch1기인대인신경효질류U251세포증식능력적영향.방법 채용Notch1-shRNA만병독감염인효질류U251세포,사선은정저표체Notch1적세포단극륭,Western blot법검측세포Notch1단백적표체;CCK-8법측정세포생장;평판극륭형성실험화연경지극륭형성실험검측세포적집락형성능력;라서충식류모형관찰Notch1침묵대충식류생장적영향.결과 성공사선득도Notch1은정침묵적U251세포단극륭,기Notch1단백하조(65.3±5.1)%.Notch1은정침묵세포적생장증식능력명현수도억제,생장곡선여대조조상비명현감만;Notch1침묵조적평판극륭형성솔위(18.6±2.5)%,저우대조조(37.0±3.3)%;Notch1침묵조연경지집락형성솔(51±5)%,저우대조조(80±4)%;Notch1침묵조라서액하충식류생장속도감만,30 d후류중(0.31±0.09)g,명현저우대조조(2.35±0.71)g;통계학분석차이균유현저성.결론 RNAi침묵Notch1기인가치인효질류U251세포적체외증식화체내성류능력하강.
Aim To investigate the effects of Notch1 knockdown by RNA interference(RNAi)on proliferation of human glioma U251 cells.Methods The human glioma U251 cells were infected with lentivirus expressing Notch1-shRNA.The knockdown effect of Notch1 in transduced U251 cells was detected by Western blot.The cell growth viability was evaluated by CCK-8 assay.Colony formation assay and soft-agar colony formation were used to measure the colony formation ability of stably transduced cells.The influence of Notch1-RNAi on the implanted tumor growth was observed.Results The Notch1-knockdown U251 cell clone was selected.Notch1 expression was down-regulated by Notch1-shRNA, and the inhibitory rate was(65.3±5.1)%.The ability of growth in Notch1-knockdown cells was decreased significantly.The plate colony formation rate of Notch1-knockdown cells was(18.6±2.5)%,whereas the rate of control cells was(37.0±3.3)%.The soft-agar colony formation rate of Notch1-knockdown cells was(51±5)%,whereas the rate of control cells was (80±4)%.Implanted glioma mouse model was successfully established.The tumor weight in Notch1-knockdown group was markedly lower than that in control group(0.31±0.09 g vs 2.35±0.71g,P<0.01).Conclusion Down-regulation of Notch1 expression by RNAi can inhibit the proliferation of human glioma U251 cells.
