DPC4/Smad4基因转染对胰腺癌细胞体外生长的抑制作用  

The inhibitory effect of DPC4 gene expression on growth of pancreatic cancer cells in vitro

作　　者：沈伟 李德春[2] 朱兴国[2] 白霞 陶国清 蔡兵 

机构地区：[1]江苏省无锡第一医院普通外科,江苏无锡214002 [2]苏州大学附属第一医院普通外科,江苏苏州215007 [3]江苏省血液研究所血栓室,江苏苏州215007

出　　处：《中国普通外科杂志》2007年第10期962-967,共6页China Journal of General Surgery

摘　　要：目的观察抑癌基因DPC4/Smad4对胰腺癌细胞生长的抑制作用。方法将人全长DPC4/Smad4cDNA亚克隆到逆转录病毒载体pLXSN中,分别导入包装细胞GP+E86和PA317,经G418筛选获取阳性抗性克隆。收集病毒上清液,转染至胰腺癌BxPC-3细胞;获稳定表达后,观察其对BxPC-3体外生长的抑制作用。结果聚合酶链反应扩增证实GP+E86、PA317/pLXSNDPC4+细胞中有外源性DPC4/Smad4基因整合;DPC4基因转染BxPC-3细胞后,可获得稳定表达,并可显著抑制胰腺癌细胞的生长和集落形成,下调癌细胞内VEGF基因的表达。结论将DPC4/Smad4逆转录病毒载体转染到人源性胰腺癌细胞后,可显著抑制癌细胞的生长及血管形成能力。Objective To observe the inhibitory effect of DPC4 gene on growth of pancreatic carcinoma in vitro. Methods The human DPC4 cDNA was subcloned to the retroviral vector pLXSN and then packaged with GP ＋ E86 and PA317 packaging cells respectively. AntiG418 clones were acquired and named as PA317/ pLXSN DPC4 ＋ cells. The DPC4 gene was restored to the human pancreatic cancer cell line BxPC-3 by the infection of the pLXSN/DPC4 and then had a stable expression after antiG418 selection, which was demonstrated by RT-PCR and Western blot. The inhibitory action of DPC4 gene expression on growth of these daughter cells was observed, Results DPC4/Smad4 gene integration in GP ＋ E86 ,PA317/ pLXSN DPC4 ＋ cells was detected by polymerase chain reaction. The BxPC-3 cells, which were null for DPC4, had stable expression of DPC4 after the infection of the pLXSN/DPC4. The expression of DPC4 gene was able to inhibit the growth of these cancer cells in vitro and downregulate the VEGF mRNA level. Conclusions This study suggested that there can be marked inhibition of growth and angiogenetic ability of pancreatic cancer cells after infection by retrovirus vector containing DPC4.

关 键 词：胰腺肿瘤 DPC4/SMAD4 逆转录病毒 基因治疗 

分 类 号：R735.9[医药卫生—肿瘤]