EP4受体拮抗剂对激素非依赖性前列腺癌DU145细胞恶性表型的影响

Effects of EP4 receptor antagonist on the malignant phenotypes of androgen-independent prostate cancer DU145 cells

目的 EP4受体与肿瘤的发生、发展关系密切,文中旨在探讨EP4受体拮抗剂对激素非依赖性前列腺癌(androgen-independent prostate cancer,AIPC)细胞的体外抗肿瘤效应。方法利用RT-PCR和Western blot检测DU145细胞中EP4受体mRNA及蛋白的表达水平。采用MTT比色法、划痕愈合实验、细胞侵袭实验观察ONO-AE3-208对AIPC DU145细胞增殖、迁移和侵袭能力的抑制效应。用已知EP4极低水平表达的LNCaP/mock和LNCaP细胞作为对照组,DU145细胞作为实验组。结果 RT-PCR及Western blot结果显示,DU145细胞中EP4受体mRNA表达水平较对照的LNCaP和LNCaP/mock细胞分别高出4.698倍和8.137倍,EP4受体蛋白也呈现高水平表达。MTT比色法结果显示随ONO-AE3-208作用时间和浓度的增加,其对DU145细胞增殖的影响不明显(P>0.05);划痕愈合实验结果提示:经10μmol/L ONO-AE3-208作用24h,DU145细胞的细胞迁移率[(16.231±4.112)%]明显低于对照组[(48.710±7.219)%](P<0.01)。细胞侵袭实验结果提示:经0.1、1.0和10.0μmol/LONO-AE3-208作用24 h,实验组DU145细胞穿入下室细胞数[分别为(27.4±4.5)、(18.6±4.0)、(13.6±5.0)个]明显少于对照组[(38.8±5.2)个],差异有统计学意义(P<0.05)。结论 EP4受体在AIPC DU145细胞中表达明显升高。EP4受体拮抗剂ONO-AE3-208对AIPC DU145的增殖无影响,对其迁移和侵袭能力有明显的抑制作用。

Objective The EP4 receptor is proved to be closely related to the development and progression of cancer.This study is to examine the anti-tumor effects of the EP4 antagonist on androgen-independent prostate cancer （AIPC） DU145 cells in vitro.Methods We detected the expression levels of EP4 mRNA and protein in the DU145 cells by quantitative real-time PCR and Westernblot,and observed the inhibitory effect of the EP4 antagonist ONO-AE3-208 on the proliferation of the AIPC DU145 cells by MTT,wound-healing test,and Transwell invasion assay,with the known extremely lowly expressed LNCaP and LNCaP/mock cells as the control.Results Quantitative real-time PCR showed that the expression level of EP4 mRNA in the DU145 cells was 4.698 and 8.137 times higher respectively than those in the LNCaP and LNCaP/mock cells,and the EP4 protein was also highly expressed.The EP4 antagonist ONO-AE3-208 exhibited no significant time-and concentration-dependent influence on the proliferation of the DU145 cells （P 〉0.05）.After treated with ONO-AE3-208 at 10 μmol/L for 24 hours,the DU145 cells showed a marked lower rate of migration than the control （[16.231 ±4.112]% vs [48.710 ±7.219]%,P 〈 0.01）.After 24-hour intervention with ONO-AE3-208 at 0.1,1.0,and 10 μmol/L,the numbers of the DU145 cells that had invaded the inferior chamber were 27.4 ±4.5,8.6 ±4.0,and 13.6 ± 5.0 respectively,significantly lower than 38.8 ± 5.2 in the control group （P 〈 0.05）.Conclusion The EP4 receptor is highly expressed in AIPC DU145 cells.The EP4 antagonist ONO-AE3-208 does not affect the proliferation of the DU145 cells,but can significantly inhibit their migration and invasion abilities.