抑制panc-1细胞的hENT1表达对氟尿嘧啶细胞毒性的影响
Impact of hENT1 gene inhibition on the cytotoxicity of 5-FU to pancreatic cancer cell line PANC-1
目的：研究用RNAi技术稳定抑制胰腺癌Panc-1细胞株平衡型核苷载体1（hENT1）的表达后，氟尿嘧啶（5-Fluorouraci，5-FU）对其细胞毒性的改变。方法：将自行设计并构建的能表达特异性针对hENT1的shRNA的重组质粒通过脂质体Lipofectamine 2000转染到胰腺癌Panc-1细胞内，通过RT-PCR鉴定干扰效率， MTT法检测干扰前后Panc-1细胞株在不同浓度5-FU(0~ 2×108 ng•L-1)中48h后的生存率，绘制生长曲线并计算各组5-FU的IC50。结果： 获得了用RNAi稳定抑制hENT1表达的Panc-1细胞株，RT-PCR结果显示干扰后的Panc-1细胞hENT1的mRNA表达水平明显下调，MTT结果显示经RNA干扰组氟尿嘧啶的IC50较未干扰组明显降低，说明对5-FU的敏感性提高。结论：抑制hENT1的表达可以提高5-FU对胰腺癌Panc-1细胞株的细胞毒性。
Objective To investigate the change of the cytotoxicity of 5-FU to PANC-1 cell line after inhibition of hENT1 expression by RNAi . Methods The pSilence-hENT1 shRNA expression vector was constructed by gene recombination, then transfected into Panc-1 cells with Lipofectamine 2000. The expression of hENT1-mRNA in Panc-1 before and after transfection was determined by RT-PCR in order to detect the efficiency of RNAi. Cells before and after RNAi were cultured respectively in the medium with a serial concentrations of 5-FU from 0 to 2×108 ng•L-1for 48 hr. Then the cell proliferation in each group was measured with MTT assay and the IC50 were evaluated. Results Obtained the Panc-1 cell line of which hENT1 expression was inhibited stably by RNAi. The mRNA expression of hENT1 was lower obviously after RNAi. The IC50 of 5-FU in RNAi group was lower obviously than in no-RNAi group. Conclusion The cytotoxicity of 5-FU to PANC-1 cell line can be increased by inhibiting the hENT1 expression.
陈武强、刘胜利
肿瘤学基础医学药学
细胞毒性胰腺癌细胞株核苷转运载体5-氟尿嘧啶RNA干扰
CytotoxicityPancreatic cancer cell lineNucleoside transporter5-FluorouraciRNAi
陈武强,刘胜利.抑制panc-1细胞的hENT1表达对氟尿嘧啶细胞毒性的影响[EB/OL].(2007-01-10)[2025-08-02].http://www.paper.edu.cn/releasepaper/content/200701-98.点此复制
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