中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2011年
5期
334-339
,共6页
刘明月%侯桂琴%张艳%贝维娟%闫爱华
劉明月%侯桂琴%張豔%貝維娟%閆愛華
류명월%후계금%장염%패유연%염애화
食管肿瘤%RNA干扰%流式细胞术%mTOR-pTOSSK信号通路%裸鼠
食管腫瘤%RNA榦擾%流式細胞術%mTOR-pTOSSK信號通路%裸鼠
식관종류%RNA간우%류식세포술%mTOR-pTOSSK신호통로%라서
Esophageal neoplasms%RNA interference%Flow cytometry%mTOR-p70S6K pathway%Nude mice
目的 检测哺乳动物雷帕霉素靶蛋白(mTOR)小分子干扰RNA(siRNA)对食管鳞癌中mTOR-p70S6K信号通路的阻断作用以及对裸鼠移植瘤生长的影响.方法 以食管鳞癌细胞株EC9706为研究对象,采用siRNA干扰、逆转录聚合酶链反应(RT-PCR)、Western blot、流式细胞术及CCK-8等方法,观察mTOR-p70S6K信号通路被阻断后,通路中各因子蛋白及mRNA表达的变化,及其对细胞增殖和凋亡的影响.进行裸鼠成瘤实验,观察mTOR siRNA对肿瘤生长的影响.结果 与未转染细胞相比,转染mTOR siRNA的细胞中mTOR和磷酸化p70S6K的表达水平降低(P<0.05),而pTOS6K的表达水平升高(P<0.05).转染mTOR siRNA后,细胞凋亡增加,细胞增殖能力降低,且对顺铂的敏感性增高,细胞被阻滞在G,期(P<0.05).mTOR siRNA能明显抑制裸鼠移植瘤的生长,siRNA组和siRNA+顺铂组的肿瘤抑制率分别为50.9%和62.3%.结论 mTOR siRNA能有效抑制mTOR-pTOS6K信号通路,抑制细胞增殖,促进细胞凋亡,在裸鼠体内能显著抑制肿瘤的生长.mTOR-pVOSSK信号通路在食管鳞癌发生发展中起重要作用.
目的 檢測哺乳動物雷帕黴素靶蛋白(mTOR)小分子榦擾RNA(siRNA)對食管鱗癌中mTOR-p70S6K信號通路的阻斷作用以及對裸鼠移植瘤生長的影響.方法 以食管鱗癌細胞株EC9706為研究對象,採用siRNA榦擾、逆轉錄聚閤酶鏈反應(RT-PCR)、Western blot、流式細胞術及CCK-8等方法,觀察mTOR-p70S6K信號通路被阻斷後,通路中各因子蛋白及mRNA錶達的變化,及其對細胞增殖和凋亡的影響.進行裸鼠成瘤實驗,觀察mTOR siRNA對腫瘤生長的影響.結果 與未轉染細胞相比,轉染mTOR siRNA的細胞中mTOR和燐痠化p70S6K的錶達水平降低(P<0.05),而pTOS6K的錶達水平升高(P<0.05).轉染mTOR siRNA後,細胞凋亡增加,細胞增殖能力降低,且對順鉑的敏感性增高,細胞被阻滯在G,期(P<0.05).mTOR siRNA能明顯抑製裸鼠移植瘤的生長,siRNA組和siRNA+順鉑組的腫瘤抑製率分彆為50.9%和62.3%.結論 mTOR siRNA能有效抑製mTOR-pTOS6K信號通路,抑製細胞增殖,促進細胞凋亡,在裸鼠體內能顯著抑製腫瘤的生長.mTOR-pVOSSK信號通路在食管鱗癌髮生髮展中起重要作用.
목적 검측포유동물뢰파매소파단백(mTOR)소분자간우RNA(siRNA)대식관린암중mTOR-p70S6K신호통로적조단작용이급대라서이식류생장적영향.방법 이식관린암세포주EC9706위연구대상,채용siRNA간우、역전록취합매련반응(RT-PCR)、Western blot、류식세포술급CCK-8등방법,관찰mTOR-p70S6K신호통로피조단후,통로중각인자단백급mRNA표체적변화,급기대세포증식화조망적영향.진행라서성류실험,관찰mTOR siRNA대종류생장적영향.결과 여미전염세포상비,전염mTOR siRNA적세포중mTOR화린산화p70S6K적표체수평강저(P<0.05),이pTOS6K적표체수평승고(P<0.05).전염mTOR siRNA후,세포조망증가,세포증식능력강저,차대순박적민감성증고,세포피조체재G,기(P<0.05).mTOR siRNA능명현억제라서이식류적생장,siRNA조화siRNA+순박조적종류억제솔분별위50.9%화62.3%.결론 mTOR siRNA능유효억제mTOR-pTOS6K신호통로,억제세포증식,촉진세포조망,재라서체내능현저억제종류적생장.mTOR-pVOSSK신호통로재식관린암발생발전중기중요작용.
Objective To investigate the effects of mTOR siRNA on mTOR-p70S6K signaling pathway in esophageal squamous cell carcinoma ( ESCC) cells in vitro, and growth and apoptosis in transplanted tumor in nude mice. Methods mTOR siRNA was transfected into ESCC cell line EC9706 cells. The expressions of factors of the mTOR/p70S6K signaling pathway were detected by RT-PCR and Western blot. DNA contents and cell apoptosis were determined by flow cytometry, and cell proliferation was measured by CCK-8 assay. The effects of mTOR siRNA on the transplanted tumor growth were assessed in nude mice. Results The levels of mTOR and p-p70S6K were significantly decreased ( P < 0.05 ) while the level of p70S6K was increased (P<0.05) in the cells transfected with mTOR siRNA, compared with that in untransfected cells and cells transfected with control siRNA. After being interfered by mTOR siRNA, the number of apoptotic cells was increased, cell proliferation became slower and cell cycle was arrested in G, phase compared with that in control cells. Also, mTOR siRNA inhibited the growth of transplanted tumor in vivo. Conclusions mTOR siRNA can effectively interfere in mTOR-p70S6K signaling pathway, induce cell apoptosis and inhibit cell proliferation and tumor growth, suggesting that mTOR-p70S6K signaling pathway plays an important role in the carcinogenesis and development of esophageal squamous cell carcinoma.
