DPC4重组质粒的构建及其对人胰腺癌细胞的抑制作用

Construction of DPC4 Gene Recombinant Expression Vector and Its Inhibitory Effects on Human Pancreatic Adenocarcinoma Cell Line Cells

目的 构建DPC4基因重组质粒以研究DPC4对人胰腺癌细胞的抑制作用。方法 应用RT PCR技术扩增出野生型DPC4基因全长cDNA ,然后用分子克隆技术构建其真核表达载体 ,应用脂质体法将DPC4基因导入到人胰腺癌PC 3细胞中 ,经G418筛选获得可稳定表达DPC4的人胰腺癌细胞 ,观察DPC4基因对人胰腺癌细胞周期和细胞增殖的影响。结果 获得了野生型DPC4真核表达质粒pcDNA3 .1 DPC4,野生型DPC4的导入可引起胰腺癌G1 期细胞的增加和S期细胞相应减少 ,同时抑制细胞生长。结论 野生型DPC4基因具有调节胰腺癌细胞增殖的功能 ,可作为胰腺癌基因治疗的靶基因。

Objective To construct DPC4 gene recombinant expression vector and to study the inhibitory effect of DPC4 on the growth of human pancreatic adenocarcinoma cell line (PC 3) cells.Methods DPC4 cDNA was amplified from K562 cell line using RT PCR, and was cloned into the pcDNA3.1 vector to construct a recombinant expression vector plasmid pcDNA3.1 DPC4. The recombinant expression plasmid was transferred into PC 3 cells by liposome method. After G418 selection, cell cycle and apoptosis were assessed by flow cytometry, then the cell growth rate was estimated by cell count. The cells being not transferred plasmid and transferred pcDNA3.1 plasmid were used as controls.Results The DPC4 gene recombinant expression vector was constructed. Wild type DPC4 gene attributed to the increase of G 1 phase cells and the decrease of S phase cells in PC 3 cells,and could inhibit the growth of PC 3 cells, the cell growth rates was reduced to 34.3%-41.1% of that of the controls, but cell apoptosis was not observed on all groups. Conclusion Wild type DPC4 gene could inhibit the proliferation of human pancreatic adenocarcinoma cells and could become one of the target genes of pancreas adenocarcinoma gene therapy.