【摘要】 目的研究Hedgehog信号通路特异性抑制剂Cyclopamine对人肺癌细胞NCI-H460生长和增殖的影响。方法培养NCI-H460细胞,加入不同浓度Cyclopamine,作用不同时间,倒置显微镜下观察细胞形态变化,CCK-8法检测Cyclopamine对肺癌细胞生长和增殖的影响,同时用流式细胞仪检测Cyclopamine对NCI-H460细胞周期的影响。结果与空白对照组和Tomatidine对照组相比,2~40μmol/L的Cyclopamine作用于NCI-H460细胞后,倒置显微镜下观察,细胞立体感消失,皱缩成圆形,细胞间隙增大。CCK-8比色法显示,Cyclopamine对NCI-H460细胞增殖的抑制作用呈剂量依赖性,20μmol/L的Cyclopamine使NCI-H460细胞生长曲线下移,并可将NCI-H460细胞阻滞在G1期,明显减少其S期细胞数量,引起细胞凋亡。结论Cyclopamine对人肺癌细胞系NCI-H460生长和增殖有比较明显的抑制作用,提示大细胞肺癌的发生和发展与Shh信号转导通路的异常激活有关。更多还原

【Abstract】 Objective To examine the effect of Cyclopamine,the inhibitor of Sonic hedgehog signal transduction pathway,on the growth and proliferation of lung cancer cell line NCI-H460.Methods NCI-H460 cells were cultured with different concentrations of Cyclopamine in vitro for different time.The inverted microscope was used to observe the morphologic changes of the cells and the CCK-8 assay and cell cycle analysis were used to examine the influence of Cyclopamine on NCI-H460 cell growth and DNA synthesis.Results Compared with blank control group and Tomatidine control group,there were obvious morphologic changes of the cells cultured with Cyclopamine at the concentrations of 2~40 μmol/L.The growth of NCI-H460 cells were inhibited by Cyclopamine in a dose-and time-dependent manner.The DNA synthesis of NCI-H460 cells was obviously inhibited,the number of NCI-H460 cells in S phase decreased and the cells were blocked in G1/S phase and cell apoptosis were found.Conclusion Cyclopamine can effectually inhibit the growth and proliferation of NCI-H460 cells by inhibiting Hedgehog signal transduction pathway.The incidence and development of large-cell lung cancer can be concerned with the abnormal activation of Hedgehog pathway.更多还原