DLL3蛋白在人小细胞肺癌细胞中的功能及其机制

为探讨DLL3(human notch ligand delta-like 3)过表达对人小细胞肺癌细胞的影响及可能的作用机制,通过PCR方法扩增人DLL3基因全长序列,并克隆至慢病毒表达载体Lenti-EFS-FLAG-puro而构建人DLL3基因过表达慢病毒表达质粒,酶切及测序鉴定质粒正确。通过慢病毒包装及感染,构建DLL3稳定过表达的人小细胞肺癌细胞株,并通过Western blot验证DLL3蛋白的表达。采用CCK-8法检测DLL3过表达对人小细胞肺癌细胞的增殖的影响。采用平板克隆实验检测DLL3过表达对人小细胞肺癌细胞的克隆形成的影响。Western blot检测DLL3过表达对细胞周期相关蛋白Cyclin D1,Cyclin D3的表达水平的影响。结果表明:人DLL3过表达慢病毒表达质粒构建成功; CCK-8实验显示DLL3过表达促进人小细胞肺癌细胞的增殖。平板克隆实验显示DLL3过表达提高人小细胞肺癌细胞的克隆形成能力。Western blot结果表明DLL3过表达增加细胞周期蛋白Cyclin D1,Cyclin D3的表达水平。可见DLL3过表达对人小细胞肺癌细胞的增殖具有促进作用。

The function and mechanism of human Notch ligand Delta-like 3 protein in human small cell lung cancer cells

Objective To construct a lentivirus expression vector overexpressing human Notch ligand Delta-like 3(DLL3) gene and investigate the effect of overexpression of DLL3 on proliferation of human small cell lung cancer cells and its possible mechanism. Methods the full-length sequence of human DLL3 gene was amplified by PCR and ligated into the lentivirus expression vector Lenti-EFS-FLAG-Puro. After verification, stable DLL3 overexpressing human small cell lung cancer cell lines were constructed by lentivirus package and infection. The expression of DLL3 protein was identified by Western blot. Proliferative activity of human small cell lung cancer cells was detected by CCK8 kit and colony formation assays. Results Human DLL3 gene overexpressing lentiviral plasmid was successfully constructed. Overexpression of DLL3 can enhance human small cell lung cancer cells proliferation of by CCK8 kit and colony formation assays. Western blot showed the expression of CylinD1 and CyclinD3 were significantly increased in DLL3 overexpression cells. Conclusion Human DLL3 gene over expressing lentiviral vector plasmid was successfully constructed. Overexpression of DLL3 can promote the proliferation of human small cell lung cancer cells.