17β-雌二醇对人甲状腺滤泡状癌WRO细胞侵袭能力的影响及其机制

Effect of 17 β-estradiol on invasion ability of human follicular thyroid carcinoma WRO cells and relevant mechanism

目的探讨17β-雌二醇(17β-estradiol,E2)对人甲状腺滤泡状癌WRO细胞侵袭能力的影响及其机制。方法用10-8mol/L的E2处理WRO细胞不同时间(0、24、48 h),倒置显微镜下观察细胞形态的变化;RT-PCR和Western blot法检测细胞中G蛋白偶联雌激素受体(G protein-coupled estrogen receptor,GPER)、CXCR1、基质金属蛋白酶2(matrix metalloproteinase2,MMP2)和MMP9基因mRNA的转录水平和蛋白的表达水平;ELISA法检测细胞中IL-8的含量;Transwell小室法检测细胞的侵袭能力。结果随着E2处理时间的延长,WRO细胞间的间隙变大,逐渐离散成单个细胞,形态呈长梭形、纺锤形;细胞中GPER、CXCR1、MMP2和MMP9基因mRNA的转录水平和蛋白的表达水平及分泌IL-8的水平均逐渐升高(P<0.05或P<0.01);细胞的侵袭能力明显增强(P<0.05)。结论 E2可通过上调GPER、CXCR1、MMP2、MMP9和IL-8的表达,促进WRO细胞的侵袭。

Objective To investigate the effect of 17 β-estradiol （E2） on invasion ability of human follicular thyroid carcinoma WRO cells as well as the relevant mechanism. Methods WRO cells were treated with 10-8 tool/L E2 for various （0, 24 and 48） hours, then observed for morphological change under inverted microscope, detet331ined for mRNA transcription levels of G protein-coupled estrogen receptor（GPER）, CXC-ehemokine receptor I（CXCR1 ） matrix metalloproteinase 2 （MMP2） and MMP9 by RT-PCR, while the protein expression levels by Western blot, the IL-8 content by ELISA, and the invasion ability by Transwell assay. Results The gaps between WRO cells increased with the increasing hours for E2 treatment until the cells were dispersed to single ones in fusifonn or spindle form. The mRNA transcription and protein expression levels of GPER, CXCR1, MMP2 and MMP9 as well as IL-8 content in the cells increased gradually（P 〈 0. 05 or P 〈 0. 01 ）, while the invasion ability of cells was enhanced significantly（P 〈 0. 05 ）. Conclusion The 17 β-estradiol promoted the invasion of WRO cells by up-regulating the expressions of GPER CXCR1, MMP2, MMP9 and IL-8.