DNA损伤诱导的长链非编码RNA(DINO)下调MMP2表达抑制人绒毛膜滋养层细胞侵袭与迁移

DNA damage induced long non-coding RNA(DINO)down-regulates the expression of MMP2 and inhibits the invasion and migration of human chorionic trophoblast cells

目的探讨DNA损伤诱导的长链非编码RNA(DINO)通过调控迁移相关因子基质金属蛋白酶2(MMP2)的表达对人绒毛膜滋养层细胞侵袭与迁移的影响。方法收集子痫前期(PE)患者和正常孕妇的胎盘组织(对照组)各20例,Masson染色观察胎盘组织结构改变,免疫荧光组织化学染色检测胎盘组织滋养层细胞MMP2的表达;实时荧光定量PCR检测DINO的表达;将HTR-8/SVneo人绒毛膜滋养层细胞转染DINO小干扰片段(si-DINO)后,实时荧光定量PCR验证DINO、MMP2 mRNA表达,Western blot法检测MMP2蛋白表达;Transwell^(TM)侵袭实验和细胞划痕实验检测细胞侵袭、迁移情况。结果与对照组相比,PE患者血压、蛋白尿水平明显增高;胎盘组织可见纤维素样坏死,绒毛膜绒毛减少;免疫荧光显示胎盘滋养层细胞MMP2表达减弱、DINO表达增加;转染si-DINO后,DINO表达降低,而MMP2 mRNA及蛋白表达增加,HTR-8/SVneo细胞侵袭和迁移能力显著增加。结论DINO下调MMP2表达,抑制胎盘滋养层细胞的侵袭与迁移。

Objective To investigate the effect of DNA damage induced long non-coding RNA(DINO)on the invasion and migration of human chorionic trophoblast cells via its regulating the expression of migration-related factor matrix metalloproteinase 2(MMP2).Methods The placental tissues of 20 patients with preeclampsia(PE)and 20 normal pregnant women(control group)were collected.The changes of placenta structure were observed by Masson staining.The expression of MMP2 in placental trophoblast cells was detected by immunofluorescence histochemical staining.The expression of DINO was detected by real-time fluorescence quantitative PCR.After DINO small interference fragments(si-DINO)were transfected into HTR-8/SVneo human chorionic trophoblast cells,the expressions of DINO and MMP2 mRNA were detected by real-time fluorescent quantitative PCR,and the expression of MMP2 protein was detected by Western blot.The invasion and migration abilities of cells were detected by Transwell^(TM) invasion assay and cell scratch assay.Results Compared withthe normal pregnant women,the PE patients’blood pressure and proteinuria levels were significantly increased;fibrinoid necrosis and chorionic villi reduction were observed in the placental tissue;immunofluorescence showed that the expression of MMP2 was significantly decreased and the expression of DINO was increased in placental trophoblast cells;after transfection with si-DINO,the expression of DINO was decreased,while the expressions of MMP2 mRNA and protein were increased,and the invasion and migration abilities of HTR-8/SVneo cells increased significantly.Conclusion DINO down-regulates the expression of MMP2 and inhibits the invasion and migration of placental trophoblast cells.