邻苯二甲酸二(2-乙基己)酯可抑制人原代培养早孕绒毛滋养细胞浸润能力

Effect of di-(2-ethylexyl) phthalate on cell invasiveness in cultured cytotrophoblasts

目的研究邻苯二甲酸二(2-乙基己)酯[di-(2-ethylexyl)phthalate,DEHP]对人原代培养早孕绒毛滋养细胞浸润能力及基质金属蛋白酶(MMP)-2、MMP-9、基质金属蛋白酶抑制因子(TIMP)-1和TIMP-2表达的影响,探讨DEHP对妊娠的影响及其毒性机制。方法2006年12月于北京军区总医院采用不同浓度DEHP预处理人原代培养早孕绒毛滋养细胞,采用Transwell体外浸润实验检测滋养细胞浸润能力的改变。RT-PCR法检测滋养层细胞侵袭性相关基因MMP-2、MMP-9、TIMP-1和TIMP-2的mRNA表达,Western blot法检测MMP-2、MMP-9、TIMP-1和TIMP-2蛋白表达水平。结果DEHP作用后,滋养细胞浸润能力明显下降。当DEEP为50、100μmol/L时,MMP-2、MMP-9、TIMP-1的mRNA表达量分别为0.35±0.07、0.26±0.03、0.97±0.18和0.30±0.10、0.20±0.05、1.02±0.20,与DEEP为0时MMP-2、MMP-9、TIMP-1的mRNA的表达量(0.77±0.15、0.45±0.04、0.80±0.20)比较P<0.05;当DEEP为50、100μmol/L时,MMP-2、MMP-9、TIMP-1的蛋白表达量分别为0.35±0.03、0.23±0.05、0.69±0.08和0.24±0.02、0.17±0.02、0.86±0.10,与DEEP为0时MMP-2、MMP-9、TIMP-1的mR-NA的表达量(0.69±0.04、0.57±0.03、0.24±0.12)比较P<0.05。可见DEHP剂量≥50μmol/L时,滋养细胞MMP-2和MMP-9表达下降、TIMP-1表达增加,且呈剂量依赖性。结论DEHP可通过抑制MMP-2、MMP-9的表达并促进TIMP-1表达,影响滋养细胞浸润能力。

Objective To investigate the influence of di-（2-ethylexyl） phthalate （DEHP） on cell invasion and expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in cultured human cytotrophoblasts. Methods After treatment of cultured cytotrophoblast with DEHP, cell matrigel invasion assay was used to assess the potential of cell invasiveness. The expression of invasiveness-associated genes, including matrix metalloproteinase （ MMP-2, MMP-9） and tissue inhibitor of metalloproteinase （TIMP-1, TIMP-2）, were detected by the method of reverse transcription polymerase chain reaction （RT- PCR） in cultured cytotrophoblast cells. The expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in eytotrophoblast cells was investigated with Western blot. Results DEHP could reduce invasiveness potential of cytotrophoblast cells by dosedependent fashion in vitro. After incubated with 501xM DEHP for 24 hours, the expression of MMP-2 and MMP-9 in cytotrophoblast cells were significantly reduced, while the expression of TIMP-1 was highly up-regulated. Conclusion DE- HP can inhibite the invasiveness of cytotrophoblast cells in a dose-dependent manner; this anti-invasive activity may be the results of decreasing MMP-2 and MMP-9 and increasing TIMP-1 expression in cytotrophoblast cell.