LncRNA-UCA1通过靶向调控miR-182-5p对滋养细胞侵袭与转移的影响

doi:10.6040/j.issn.1671-7554.0.2021.0660

摘要/Abstract

摘要： 目的探讨长链非编码RNA尿道上皮癌相关基因1(LncRNA-UCA1)对滋养细胞侵袭与转移的影响及其可能机制。方法收集30例行剖宫产分娩的子痫前期(PE)患者胎盘组织和30例正常妊娠孕妇胎盘组织,qRT-PCR检测各胎盘组织中UCA1和miR-182-5p表达水平。以滋养细胞HTR-8/Svneo为研究对象,将UCA1过表达质粒(pcDNA3.1-UCA1)及其空载质粒(Vector)转染至细胞中,qRT-PCR检测细胞中UCA1和miR-182-5p表达水平;采用双荧光素酶报告基因实验验证UCA1与miR-182-5p之间的靶向调控关系。将pcDNA3.1-UCA1质粒与miR-182-5p mimic分别或同时转染至HTR-8/Svneo细胞中,CCK-8检测细胞增殖活性;Transwell实验检测细胞侵袭与转移;Western blotting检测基质金属蛋白酶-2(MMP-2)和MMP-9蛋白表达变化。结果 (1)临床标本测定结果:与正常妊娠孕妇胎盘组织比较,PE患者胎盘组织中UCA1表达降低(1.10±0.51 vs 5.74±1.63),差异有统计学意义(t =14.947,P<0.001),而miR-182-5p表达显著增加(7.23±1.44 vs 1.07±0.72),差异有统计学意义(t=20.961,P<0.001)。(2)细胞实验结果:UCA1能特异性靶向调控miR-182-5p表达。与Vector组比较,UCA1过表达可降低HTR-8/SVneo细胞中miR-182-5p表达(0.12±0.05 vs 0.97±0.06,t=19.028,P<0.001),促进细胞增殖、侵袭和迁移(t=10.160、17.228、13.768,P均<0.001),并上调MMP-2和MMP-9蛋白表达(t=20.833、24.793,P均<0.001);而miR-182-5p过表达能显著减弱UCA1过表达对HTR-8/SVneo细胞增殖、侵袭和迁移的促进作用。结论 UCA1通过靶向下调miR-182-5p表达促进滋养细胞侵袭与转移。

关键词: 长链非编码RNA, 尿道上皮癌相关基因1, 子痫前期, 滋养细胞, 侵袭, 转移

Abstract: Objective To investigate the effect of long non-coding RNA urethelial carcinoma related gene 1(LncRNA-UCA1)on invasion and metastasis of trophoblast cells and its possible mechanism. Methods Thirty placentas tissues from patients with preeclampsia(PE)and other 30 normal placentas tissues were collected respectively. The expression levels of UCA1 and miR-182-5p in placentas tissues were detected by qRT-PCR. UCA1 overexpression plasmid(pcDNA3.1-UCA1)and its negative plasmid(Vector)were transfected into HTR-8/Svneo cells respectively, and the expression levels of UCA1 and miR-182-5p in cells were detected by qRT-PCR. The targeted regulatory relationship between UCA1 and miR-182-5p was verified by dual-luciferase reporter assay. HTR-8/Svneo cells were transfected with pcDNA3.1-UCA1 plasmid and miR-182-5p mimic respectively or simultaneously, and then cell proliferation activity was detected by CCK-8. Cell invasion and metastasis was detected by Transwell assay. The expression levels of matrix metalloproteinase(MMP)-2 and MMP-9 proteins were detected by Western blotting. Results (1) Results of clinical specimen determination: compared with the placental tissues of normal pregnant women, the expression of UCA1 in placental tissues of PE patients was significantly decreased(1.10±0.51 vs 5.74±1.63, t=14.947, P<0.001), while the expression of miR-182-5p was significantly increased(7.23±1.44 vs 1.07±0.72, t=20.961,P<0.001). (2) Results of cell experiment: UCA1 could specifically target and regulate miR-182-5p expression. Compared with the Vector group, overexpression of UCA1 decreased the expression level of miR-182-5p in HTR-8/SVneo cells(0.12±0.05 vs 0.97±0.06, t=19.028, P<0.001), promoted the proliferation, invasion and migration of HTR-8/SVneo cells(t=10.160, 17.228, 13.768, all P<0.001), and up-regulated the expressions of MMP-2 and MMP-9 protein(t=20.833, 24.793, both P<0.001). However, the overexpression of miR-182-5p significantly reduced the promotion of UCA1 overexpression on the proliferation, invasion and migration of HTR-8/SVneo cells. Conclusion UCA1 promotes trophoblast invasion and metastasis by targeting the down-regulation of miR-182-5p.

Key words: Long non-coding RNA, Urethelial carcinoma related gene 1, Preeclampsia, Trophoblast cell, Invasion, Metastasis

中图分类号:

R714

钟黎黎,盛莹,郭江虹,阳双健,何宜静. LncRNA-UCA1通过靶向调控miR-182-5p对滋养细胞侵袭与转移的影响[J]. 山东大学学报 (医学版), 2022, 60(3): 76-82.

ZHONG Lili, SHENG Ying, GUO Jianghong, YANG Shuangjian, HE Yijing. LncRNA-UCA1 effects invasion and metastasis of trophoblast cells by targeting miR-182-5p[J]. Journal of Shandong University (Health Sciences), 2022, 60(3): 76-82.

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