Enhancement of gemcitabine sensitivity in pancreatic cancer by co-regulation of dCK and p8 expression
Enhancement of gemcitabine sensitivity in pancreatic cancer by co-regulation of dCK and p8 expression
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通过共同调节 dCK 和 p8 表达增强胰腺癌中吉西他滨的敏感性
DOI：
10.3892/or.2011.1139
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发表时间：
2011-04-01
期刊：
影响因子：
4.2
通讯作者：
Wang, Yu
中科院分区：
文献类型：
--
作者：
Tang, Ke;Zhang, Zhongtao;Wang, Yu
本研究的目的是通过腺病毒介导的dCK和p8表达的共调节来提高胰腺癌吉西他滨的敏感性。首先，我们用MTT法分析了三种人胰腺癌细胞系（Capan-2，Panc-1和BxPc-3）对吉西他滨的敏感性，发现Capan-1对吉西他滨相对耐药。此外，我们还利用实时荧光定量PCR和Western blot分析了dCK和p8在不同胰腺癌细胞系中的表达，发现这两个基因的表达水平与胰腺癌细胞对吉西他滨的敏感性有关。我们构建了重组腺病毒载体Ad-dCK和Ad-p8-siRNA，分别过表达dCK和敲低p8。使用MTT法，我们观察到使用Ad-dCK和Ad-p8-siRNA在体外联合感染导致吉西他滨IC 50显著降低，并且在对吉西他滨相对耐药的NSCLC-1细胞中凋亡和caspase-3活性增加。此外，在建立的裸鼠皮下胰腺癌模型中，在腹腔内吉西他滨化疗的基础上，瘤内注射Ad-dCK和Ad-p8-siRNA后，肿瘤抑制显著增强，并伴随着凋亡指数的升高。综上所述，本研究结果表明，dCK和p8可能是调节胰腺癌细胞吉西他滨敏感性的重要因素。此外，这两个因素的共同调节比单独调节效果更好。
The purpose of this study was to improve the gemcitabine sensitivity in pancreatic cancer by adenovirus-mediated co-regulation of dCK and p8 expression. Firstly, we analyzed the sensitivity of three human pancreatic tumor cell lines (Capan-2, Panc-1 and BxPc-3) to gemcitabine using MTT assays, and found Pane-1 to be relatively resistant to gemcitabine. Further, we investigated the expression of dCK and p8 in different pancreatic cancer cell lines using real-time PCR and Western blot analysis, and found that the expression levels of these two genes were related to the gemcitabine sensitivity of pancreatic cancer cells. We constructed recombinant adenovirus vectors, Ad-dCK and Ad-p8-siRNA, that overexpressed dCK and knocked down p8, respectively. Using MTT assays, we observed that combined infection using Ad-dCK and Ad-p8-siRNA in vitro led to a significant decrease in the gemcitabine IC50 with an increase in apoptosis and caspase-3 activity in Pane-1 cells, which are relatively resistant to gemcitabine. Furthermore, in established subcutaneous pancreatic cancer models in nude mice, the tumor inhibition was markedly enhanced accompanied by elevation of the apoptosis index after intratumoral injection of Ad-dCK and Ad-p8-siRNA on the basis of intraperitoneal gemcitabine chemotherapy. Taken together, the present findings suggest that, dCK and p8 may be the important factors in the regulation of gemcitabine sensitivity in pancreatic cancer cells. Moreover, co-regulation of the two factors achieved better effects than regulation of either one alone.