中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2011年
10期
604-607
,共4页
癌,鳞状细胞%细胞周期%RNA干扰
癌,鱗狀細胞%細胞週期%RNA榦擾
암,린상세포%세포주기%RNA간우
Carcinoma,squamous cell%Cell cycle%RNA interference
目的 应用RNA干扰技术抑制人舌鳞状细胞癌细胞株Tca8113中双泛素(diubiquitin)的表达,探讨双泛素表达下调对舌鳞状细胞癌细胞生物学行为的影响.方法 构建针对双泛素特异性小干扰RNA( small interfering RNA,siRNA)真核表达载体pU-双泛素-siRNA,将其转染至Tca8113细胞,采用反转录聚合酶链反应(RT-PCR)、蛋白质印迹法检测转染后的Tca8113细胞中双泛素的表达.通过流式细胞仪分析siRNA对舌鳞状细胞癌细胞周期的影响,并通过细胞侵袭能力实验观察siRNA对Tca8113细胞恶性生物学行为的变化.结果 siRNA干扰Tca8113细胞后,双泛素的表达无论是在mRNA水平还是在蛋白质水平均显著下降[mRNA:实验组(0.36±0.03)、空白对照组(0.92±0.07)、空载体组(0.95±0.05)；蛋白:实验组(0.39±0.04)、空白对照组(0.64±0.05)、空载体组(0.69±0.05)](P＜0.05),细胞周期被阻滞在G1期,细胞生长缓慢,体外侵袭能力下降(P＜0.05).结论 通过RNAi技术阻断双泛素的表达,可抑制Tca8113细胞的生长、增殖、侵袭,提示双泛素在舌鳞状细胞癌的发生、发展过程中起重要作用.
目的 應用RNA榦擾技術抑製人舌鱗狀細胞癌細胞株Tca8113中雙汎素(diubiquitin)的錶達,探討雙汎素錶達下調對舌鱗狀細胞癌細胞生物學行為的影響.方法 構建針對雙汎素特異性小榦擾RNA( small interfering RNA,siRNA)真覈錶達載體pU-雙汎素-siRNA,將其轉染至Tca8113細胞,採用反轉錄聚閤酶鏈反應(RT-PCR)、蛋白質印跡法檢測轉染後的Tca8113細胞中雙汎素的錶達.通過流式細胞儀分析siRNA對舌鱗狀細胞癌細胞週期的影響,併通過細胞侵襲能力實驗觀察siRNA對Tca8113細胞噁性生物學行為的變化.結果 siRNA榦擾Tca8113細胞後,雙汎素的錶達無論是在mRNA水平還是在蛋白質水平均顯著下降[mRNA:實驗組(0.36±0.03)、空白對照組(0.92±0.07)、空載體組(0.95±0.05)；蛋白:實驗組(0.39±0.04)、空白對照組(0.64±0.05)、空載體組(0.69±0.05)](P＜0.05),細胞週期被阻滯在G1期,細胞生長緩慢,體外侵襲能力下降(P＜0.05).結論 通過RNAi技術阻斷雙汎素的錶達,可抑製Tca8113細胞的生長、增殖、侵襲,提示雙汎素在舌鱗狀細胞癌的髮生、髮展過程中起重要作用.
목적 응용RNA간우기술억제인설린상세포암세포주Tca8113중쌍범소(diubiquitin)적표체,탐토쌍범소표체하조대설린상세포암세포생물학행위적영향.방법 구건침대쌍범소특이성소간우RNA( small interfering RNA,siRNA)진핵표체재체pU-쌍범소-siRNA,장기전염지Tca8113세포,채용반전록취합매련반응(RT-PCR)、단백질인적법검측전염후적Tca8113세포중쌍범소적표체.통과류식세포의분석siRNA대설린상세포암세포주기적영향,병통과세포침습능력실험관찰siRNA대Tca8113세포악성생물학행위적변화.결과 siRNA간우Tca8113세포후,쌍범소적표체무론시재mRNA수평환시재단백질수평균현저하강[mRNA:실험조(0.36±0.03)、공백대조조(0.92±0.07)、공재체조(0.95±0.05)；단백:실험조(0.39±0.04)、공백대조조(0.64±0.05)、공재체조(0.69±0.05)](P＜0.05),세포주기피조체재G1기,세포생장완만,체외침습능력하강(P＜0.05).결론 통과RNAi기술조단쌍범소적표체,가억제Tca8113세포적생장、증식、침습,제시쌍범소재설린상세포암적발생、발전과정중기중요작용.
Objective To study the effect of diubiquitin (FAT10) down-regulation by small interfering RNA-mediated RNA interference (RNAi) on the biological features of tongue carcinoma cell line Tca8113.Methods Tca8113 cells were transfected with synthetic small interfering RNA(siRNA) targeting FAT10.Expression of FAT10 mRNA and protein were respectively measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting,transfection efficiencies were monitored.The distribution of cell cycle phases was determined using flow cytometry.The proliferative and invasive ability of Tca8113 cells in vitro was evaluated by the colony-forming unit assay and Transwell migration assay respectively.Results Both FAT10 mRNA and protein expression were significantly decreased in the experimental group ( pU-FAT10-siRNA:mRAN 0.36 ± 0.03,Protein 0.39 ± 0.04) compared with controls ( Control:mRNA 0.95 ± 0.05,Protein 0.69 ± 0.05 ; pU-siRNA:mRNA 0.92 ± 0.07,Protein 0.64 ± 0.05 ) (P ＜ 0.05 ).The cell cycle was arrested in the G1 phase [ pU-FAT10-siRNA:(72.45 ± 5.81 ) ％,Control:(45.95 ± 3.80 ) ％,pU-siRNA:(45.95 + 3.80) ％ ].The proliferation and invasiveness of treated Tca8113 cells were inhibited in vitro ( pU-FAT10-siRNA:41.83 ± 8.19,Control:317.21 ± 69.48,pU-siRNA:339.36 +73.84).Conclusions Delivery of siRNA targeting FAT10 seems efficient in down-regulating FAT10 expression and diminishing the growth,proliferation and invasiveness of Tca8113 cells,suggesting that siRNA-based strategy targeting FAT10 may lay a foundation for the clinical management of tongue carcinoma.
