miR-23c对滋养层细胞增殖、侵袭及迁移的影响及作用机制研究

Effects and mechanism of miR-23c on proliferation,invasion and migration of trophoblast cells

目的探讨miR-23c对滋养层细胞HTR8/SVneo增殖、侵袭和迁移的影响及作用机制。方法选取2019年4月至2020年8月在北海市人民医院行产前检查的子痫前期(PE)及正常妊娠孕妇各35例,收集血清及胎盘组织,采用qRT-PCR法检测miR-23c表达。将HTR8/SVneo细胞分为Blank control组、mimic-NC组、miR-23c mimic组、inhibitor-NC组和miR-23c inhibitor组,采用qRT-PCR法及Western blot法分别检测miR-23c及磷酸酶和张力蛋白同源物(PTEN)、蛋白激酶B(Akt)mRNA和蛋白及p-Akt表达;CCK-8法、细胞划痕实验和Transwell实验分别检测细胞的增殖、侵袭及迁移能力;双荧光素酶报告实验验证miR-23c对PTEN的靶向作用。结果与正常孕妇相比,PE患者血清及胎盘组织中miR-23c表达水平均显著降低(均P<0.05)。与Blank control组相比,miR-23c mimic组miR-23c、p-Akt/Akt表达水平均显著升高(均P<0.05),PTEN表达水平显著降低(P<0.05),细胞增殖、侵袭及迁移能力均显著增强(均P<0.05);miR-23c inhibitor组miR-23c、p-Akt/Akt表达水平均显著降低(均P<0.05),PTEN表达水平显著升高(P<0.05),细胞增殖、侵袭及迁移能力显著减弱(均P<0.05)。Blank control组、mimic-NC组和inhibitor-NC组以上各项指标比较差异无统计学意义(P>0.05)。双荧光素酶报告实验显示miR-23c可靶向抑制PTEN的表达。结论miR-23c通过靶向抑制PTEN的表达来促进Akt活化,进而促进滋养层细胞的增殖、侵袭和迁移。

Objective To investigate the effects and mechanism of miR-23c on the proliferation,invasion and migration of trophoblast HTR8/SVneo cells.Methods Serum and placental tissue samples were collected from 35 preeclampsia(PE)patients and 35 normal pregnant women,and the expression of miR-23c was detected by qRT-PCR.Human trophoblast HTR8/SVneo cells were divided into blank control group,mimic-NC group,miR-23c mimic group,inhibitor-NC group and miR-23c inhibitor group.The expression levels of miR-23c,Akt,p-Akt and PTEN were detected by qRT-PCR and Western blotting.CCk-8 assay,cell scratch assay and Transwell assay were used to observe cell proliferation,invasion and migration ability.The direct targeting relationship of miR-23c to PTENwas confirmed by a double luciferase reporter assay.Results Compared with the normal group,the expression levels of miR-23c in serum and placental tissues of PE patients were significantly decreased(P<0.05).Compared with the blank control group,the expression levels of miR-23c and p-Akt/Akt were significantly increased,while the expression level of PTEN was significantly decreased,and cell proliferation,invasion and migration were significantly enhanced in the miR-23c mimic group(P<0.05).Compared with the blank control group,the expression levels of miR-23c and p-Akt/Akt were significantly decreased,while the expression level of PTEN was significantly increased,and cell proliferation,invasion and migration were significantly decreased in the miR-23c inhibitor group(P<0.05).There was no significant difference in the detection results of blank control group,mimic-NC group and inhibitor-NC group(P>0.05).Conclusion MiR-23c promotes the activation of Akt through targeted PTEN,thus promoting the proliferation,invasion and migration of trophoblast cells.