EP4对人甲状腺乳头状癌TPC-1细胞生长的影响

Effect of EP4 on the Growth of Human Papillary Thyroid Carcinoma Cell Line TPC-1

【目的】探讨前列腺素E受体4(EP4)对人甲状腺乳头状癌TPC-1细胞生长的影响。【方法】体外培养人甲状腺乳头状癌细胞株TPC-1及人甲状腺滤泡上皮细胞株Nthy-ori 3-1,Nthy-ori 3-1细胞作为对照,用ELISA法检测细胞上清液中前列腺素E2(PGE2)含量;用实时荧光定量PCR法检测四种前列腺素E2受体亚型(EP1、EP2、EP3、EP4)m RNA表达水平;用Western Blot法检测EP1-4、PKA及PI3K两种亚型(p110α、p110γ)蛋白的表达情况。用噻唑蓝(MTT)比色法检测EP4受体拮抗剂L-161982对两种细胞生长的作用。【结果】两种细胞均分泌PGE2。与Nthy-ori 3-1细胞相比,TPC-1细胞EP2、EP4表达显著上调(P<0.05),EP4下游信号因子PI3K亚型p110α、p110γ蛋白表达升高(P<0.05)。L-161982呈浓度依赖性抑制TPC-1细胞生长(P<0.05)。【结论】EP4受体拮抗剂L-161982可抑制TPC-1细胞生长。

[Objective] To explore the effect of prostaglandin E receptor 4 （EP4） on the growth of human papillapy thyroid carcinoma cell line TPC-1. [Methods] Human papillary thyroid carcinoma cell line TPC-1 and human thyroid follicular epithelial cell line Nthy-ori 3-1 were cultured. ELISA assay was used to measure the level of PGE2 in cell supernatants. The mRNA expression of four EP receptor subtypes （EP1, EP2, EP3 and EP4） was examined by quantitative real-time PCR （qRT-PCR）. The levels of EPI-4, PKA and PI3K（p110（1 and p110γ） protein were determined by Western blotting. The effects of selective EP4 antagonist L-161982 on cell viability were determined using MTI＇ assay. [Results] PGE2 production ean be detected in both TPC-1 and Nthy-ori 3-1 cells. EP2 and EP4 expression appeared to increase in TPC-1 cells compared with Nthy-ori 3-1 cells（P 〈 0.05）. Meanwhile, TPC-1 cells expressed elevated protein levels for PI3K isoforms p 110α and p110γ in comparison withNthy-ori 3-1 cells （P 〈 0.05）.L-161982 decreased TPC-I cell viability in a dose-dependent manner（P 〈 0.05）. [Conclusion] EP4 antagonist L-161982 can inhibit the growth of TPC- 1 cells.