中文摘要：

目的：应用intrakine技术构建CXCR4（chemokine receptor 4趋化因子受体4）敲减表型的肺癌细胞,体外观察其增殖、凋亡、侵袭、转移能力的变化。方法：经克隆PCR、测序鉴定质粒后,给高表达CXCR4、高骨转移细胞系SBC-5中转染pCMV-S-K,经流式及细胞免疫化学检测成功构建了CXCR4稳定敲减表型的肺癌细胞SBC-5/S-K,检测CXCR4敲减后细胞增殖、凋亡、侵袭及转移能力的变化。结果：与对照SBC-5/neo相比,SBC-5/S-K在增殖、克隆形成、凋亡及细胞进程方面没有变化,而侵袭、转移能力明显减弱。结论：体外实验证实CXCR4是肺癌转移基因,为后续体内功能研究打下基础。

英文摘要：

Objective：To successfully produce a knock-out phenotype of CXCR4 in SBC-5 which has high potential of bone metastasis and high CXCR4 expression,the affect on cell proliferation apoptosis invasion and metastasis in vitro are researched.Methods：The pCMV-S-K vector was identified by colony PCR and DNA sequencing,then the pCMV-S-K and mock vector pCMV-neo as a negative control were transfected into SBC-5 and acquired the stable transfected SBC-5 / S-K and SBC-5 / neo cells with limiting dilution.Flow cytometric analysis was used to observe the downregulation of CXCR4 expression and immunofluorescence was used to observe the location of SDF-1 protein in SBC-5 cells.Then the affect on cell proliferation apoptosis invasion and metastasis in vitro were researched by MTT flow cytometry and transwell analysis.Results：The vector pCMV-S-K significantly decreased CXCR4 expression on SBC-5 cells compared with control.Down regulation of CXCR4 expression can reduce the cell migration and invasion,but does not affect the proliferation and apoptosis in vitro.Conclusion：CXCR4 is a candidate tumor metastasis gene of SCLC,which gaves a way for continuous studying of CXCR4 function and gene therapy in vivo.