摘要
目的探讨不同分化程度的食管鳞癌细胞对雷帕霉素的敏感性差异以及以p70S6K—siRNA干扰p70S6K表达后食管鳞癌EC9706细胞对雷帕霉素敏感性的变化。方法以细胞计数盒8（CCK-8）法检测雷帕霉素对不同分化程度食管鳞癌细胞（EC9706、TE-1、Eca109、KYSE790、KYSE450）增殖的影响，并根据检测结果，选取对雷帕霉素不敏感的EC9706细胞株转染p70S6K—siRNA。采用CCK-8法、流式细胞术和裸鼠成瘤实验，检测转染前后EC9706细胞对雷帕霉素敏感性的变化。结果CCK-8法检测结果显示，5株食管鳞癌细胞对低浓度雷帕霉素（≤100nmol／L）均敏感，而低分化细胞株TE-1和EC9706对高浓度雷帕霉素却易产生耐药性。经50、100、200、500和1000nmol／L雷帕霉素＋p70S6K—siRNA处理后，EC9706细胞的增殖率分别为（48．67±1.68）％、（15．45±1．54）％、（14．00±0．91）％、（10．97±0．72）％和（2．70±0．32）％，均分别明显低于50、100、200、500和1000nmol／L雷帕霉素＋对照siRNA处理组[（74．53±1．71）％、（68．27±1．35）％、（71．74±2．44）％、（76．23±1．02）％和（80．21±2．77）％，均P〈0．05]。流式细胞仪检测结果显示，p70S6K—siRNA组、雷帕霉素组和pTOS6K-siRNA＋雷帕霉素组EC9706细胞中G1期细胞的比例分别为（53．82±1．78）％、（57．87±4．01）％和（73．73±3．68）％，均明显高于对照组[（46．09±2．31）％，均P〈0．05]。裸鼠体内实验的结果显示，转染p70S6K-siRNA后，雷帕霉素对裸鼠体内移植瘤生长的抑制作用也增强，抑瘤率高达96．5％。结论不同分化程度的食管鳞癌细胞对雷帕霉素的敏感性不同，p70S6K-siRNA在体内外均能提高食管鳞癌EC9706细胞对雷帕霉素的敏感性。
Objective To explore the differences in sensitivity to rapamycin of five esophageal squamous cell carcinoma cell lines with different differentiation and the changes of sensitivity of the cells after siRNA-interfered expression of p70S6K. Methods Effects of rapamycin on proliferation of ESCC cell lines with different differentiation, EC9706, TE-1, Eca109, KYSE790 and KYSE450 cells, were investigated using cell counting kit 8 （CCK-8） assay, and according to the above results, the EC9706 cells non-sensitive to rapamycin were chosen to be transfected with pTOS6K-siRNA. The changes in sensitivity of cells to rapamycin were measured in vitro and in vivo using CCK-8 kit, flow cytometry and tumor formation in nude mice. Results CCK-8 results showed that all the five cell line ceils were sensitive to low concentration of rapamycin （〈 100 nmol/L）, but TE-1 and EC9706 cells, which were with poor differentiation, showed resistance to high concentration of rapamycin. After EC9706 cells were treated with 50, 100, 200, 500 and 1 000 nmol/L rapamyein and p70S6K-siRNA, the proliferation rates of EC9706 ceils were （48.67± 1.68）%, （15.45± 1.54）%, （14.00±0.91）%, （10.97±0.72）% and （2.70±0.32）%, respectively, and were significantly lower than that of cells treated with 50, 100, 200, 500 and 1 000 nmol/L rapamyein and control siRNA [ （74.53±1.71）%, （68.27±1.35）%, （71.74±2.44）%, （76.23±1,02）% and （80.21±2.77）%] （P〈0.05 for all）. The results of flow eytometry showed that the ratios of cells in G1 phase of the p70S6K- siRNA, rapamycin and p70S6K-siRNA＋rapamycin groups were （ 53.82± 1.78 ） %, （ 57.87±4.01 ） % and （73.73±3.68） %, respectively, significantly higher than that in the control group （46.09±2.31 ） % （ P〈0.05 for all）. The results of tumor formation test in vivo showed that the inhibitory effect of rapamycin on tumor growth was stronger after the cells were transfected with p70S6K-siRNA, and the inhibition rate was 96.5%. Conclusion ESCC cells with different differentiation have different sensitivity to rapamycin, and p70S6K- siRNA can improve the sensitivity of cells to rapamycin in vitro and in vivo.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2015年第12期885-889,共5页
Chinese Journal of Oncology
基金
国家自然科学基金（30901778）
郑州大学研究生教育科研专项基金
