畸胎瘤细胞源性生长因子shRNA增强胰腺癌细胞株Panc-1对健择敏感性的研究

Inhibition of PC cell-derived growth factor expression by shRNA transfection increases sensitivity of pancreatic cancer cell lines panc-1 to gemcitabine

目的通过转染针对畸胎瘤细胞源性生长因子（PCDGF）shRNA至胰腺癌细胞株Panc-1中，观察对健择敏感性的变化，分析PCDGF因子对化疗药物敏感性的影响。方法免疫组织化学和定量聚合酶链反应（PCR）测定PCDGF在胰腺癌组织中的表达，构建含2条shRNA的RNAi质粒，脂质体转染PCDGFshRNA至Panc-1，测定转染前后细胞对健择敏感性的变化。结果PCDGF表达于多数胰腺癌细胞的细胞质中（阳性率为84．5％），显示胰腺癌组织PCDGF的表达明显高于胰腺囊腺瘤和胰腺炎（t=11．41，P〈0．05），构建PCDGFshRNA质粒经脂质体转染后PC—DGF的表达显著受到抑制（抑制率为82．3％），在较高浓度时，健择的细胞毒作用明显增强。结论针对PCDGF的shRNA能显著增加Panc-1对健择的敏感性。

Objective To detect the effects of PC cell-derived growth factor （PCDGF） on sensitivity of pancreatic cancer cell lines Panc-1 to gemcitabine by observing the changes after transfection. Methods The expression of PCDGF was detected in pancreatic cancer tissues and noncarcinoma tissues by using immunohistochemical staining and quantitative reverse transcription PCR （t = 11.41 ,P 〈 0.05 ）. Two sites of shRNA targeted to PCDGF were combined to pGenesil-1 vector, and transfected to Panc-1 with lipofectamine2000, and the sensitivity of cancer cell lines to gemcitabine was measured. Results The expression level of PCDGF in pancreatic cancer tissues was higher than in pancreatic cytoadenoma and necrosis tissues in pancreatitis. Transfection of shRNA targeted to PCDGF into Panc-1 ceils could inhibit the PCDGF expression （inhibition rate = 82.3 % ）. After transfection, the toxicity of gemcitabine to Panc-1 was remarkably increased especially in high concentration. Conclusion shRNA targeting PCDGF makes pancreatic cancer cell lines Pane-1 more sense to gemcitabine.