Cyclopamine对人肺癌细胞系NCI-H460生长和增殖的影响

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。

Effect of Cyclopamine on growth and proliferation of lung cancer cell line NCI-H460

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.