化疗药顺铂对Wnt通路抑制因子的表达作用

Effects of antitumor-drug cisplatin on expression of Wnt signaling pathway inhibitor

目的研究抗肿瘤药顺铂(cisplatin,Pt)对Wnt通路抑制因子FrpHE(frizzled relatd protein)和DKK-1(dickkopf-1)表达的调节作用。方法培养人肝癌HepG2、Hep3B、人大肠癌Lovo和人神经胶质瘤U251细胞,并分别加入5μmol/L Pt作用24h,以RT-PCR技术检测FrpHE和DKK-1mRNA,以流式细胞术检测肿瘤细胞中Wnt通路的关键调节因子β-catenin的表达。结果顺铂作用24h后,FrpHE mRNA表达水平在人肝癌细胞较对照组表达水平显著增加(P<0.001)。在人大肠癌细胞和人神经胶质瘤细胞中,未见FrpHE mRNA表达。DKK-1mRNA表达水平在加入Pt作用后的人肝癌细胞、人大肠癌细胞和人神经胶质瘤细胞均较对照组表达水平显著增加。β-catenin的阳性细胞百分比和平均荧光强度与对照组相比均降低(P<0.001)。结论化疗药顺铂能够诱导肝癌细胞FrpHE mRNA和人肝癌、肠癌、神经胶质瘤细胞DKK-1mRNA的表达,抑制Wnt通路。

Objective To investigate the effects of antitumor-drug cisplatin on the expression of secreted frizzled-related protein （sFRP） and Dickkopf-1 （DKK-1）, a Wnt pathway inhibitor. Methods Human Hep3B, HepG2, Lovo, and U251 cells were transfected with antitumor-drug cisplatin, -catenin expression was measured by fluorescence-activated cell sorting （FACS） and then wnt signaling pathway inhibitor FrpHE and DKK-1 expression was detected by （ RT）-PCR. Results FrpHE mRNA in Hep3B and HepG2 cells was initially potentiated by antitumor-drug cisplatin after 24 h, no inhibitor FrpHE was expressed in Lovo and U251, DKK-1 mRNA in Hep3B, HepG2, Lovo, and U251 cells was initially potentiated by antitumor-drug cisplatin after 24 h,β-catenin positive ceils percent and fluorescence intensity were descent. Conclusion Antitumor-drug cisplatin promotes mRNA expression of wnt signaling pathway inhibitor DKK-1 and FrpHE, and Wnt pathway was inhibited.