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罗哌卡因通过调控长链非编码RNA OGFRP1/微小RNA-4731-5p/缺氧诱导因子3A轴来抑制绒毛膜癌细胞的活力、迁移和侵袭。

Ropivacaine Retards the Viability, Migration, and Invasion of Choriocarcinoma Cells by Regulating the Long Noncoding RNA OGFRP1/MicroRNA-4731-5p/HIF3A Axis.

作者信息

Lu Yaojun, Yang Chen, Zhang Le, Ding Juan

机构信息

Department of Anaesthesiology, The Obstetrics and Gynecology Hospital of Fudan University, No. 128, Shenyang Road, Shanghai City, 200090, China.

Department of Anaesthesiology, Fudan University Shanghai Cancer Center, No. 270, Dong'an Road, Shanghai City, 200032, China.

出版信息

Mol Biotechnol. 2022 May;64(5):499-509. doi: 10.1007/s12033-021-00429-1. Epub 2021 Dec 2.

Abstract

Choriocarcinoma is an aggressive gestational trophoblastic neoplasm. This study attempted to explore the biological functions and underlying mechanisms by which ropivacaine restrains the progression of choriocarcinoma. The expression of long noncoding RNA OGFRP1, microRNA-4731-5p (miR-4731-5p), and HIF3A in choriocarcinoma cells was assessed by qRT-PCR. Choriocarcinoma cells treated with ropivacaine at the concentration of 100, 500, and 1000 μM were cultured for 24, 48, and 72 h, respectively. Choriocarcinoma cell viability was evaluated by MTT assay. Transwell assay was conducted to examine choriocarcinoma cell migration and invasion. Additionally, the target relationship between OGFRP1 and miR-4731-5p or between miR-4731-5p and HIF3A was predicted by bioinformatics analysis and confirmed by dual-luciferase reporter assays. OGFRP1 and HIF3A expression were enhanced in choriocarcinoma cells, while miR-4731-5p expression was inhibited. Treatment with ropivacaine impeded choriocarcinoma cell viability, migration, and invasion. Choriocarcinoma cells treated with 1000 μM ropivacaine for 48 h were selected for subsequent experiments. OGFRP1 elevation or miR-4731-5p deficiency mitigated the reduction effect of ropivacaine on tumorigenesis of choriocarcinoma cells. Besides, miR-4731-5p was predicted as the potential OGFRP1 target by StarBase and LncBase, and HIF3A was predicted as the potential miR-4731-5p target by StarBase and TargetScan. Dual-luciferase reporter assays determined that miR-4731-5p was a target of OGFRP1 and HIF3A was a target of miR-4731-5p. Feedback experiments declared that miR-4731-5p elevation or HIF3A suppression reversed the promoting effect of OGFRP1 overexpression on the malignant behaviors of ropivacaine-treated choriocarcinoma cells. Ropivacaine constrained choriocarcinoma cell viability, migration, and invasion through modulating the OGFRP1/miR-4731-5p/HIF3A axis. Our study may provide a novel strategy for choriocarcinoma prevention and treatment.

摘要

绒毛膜癌是一种侵袭性妊娠滋养层肿瘤。本研究试图探索罗哌卡因抑制绒毛膜癌进展的生物学功能及潜在机制。通过qRT-PCR评估绒毛膜癌细胞中长链非编码RNA OGFRP1、微小RNA-4731-5p(miR-4731-5p)和HIF3A的表达。分别用浓度为100、500和1000μM的罗哌卡因处理绒毛膜癌细胞24、48和72小时。通过MTT法评估绒毛膜癌细胞活力。进行Transwell实验以检测绒毛膜癌细胞的迁移和侵袭能力。此外,通过生物信息学分析预测OGFRP1与miR-4731-5p之间或miR-4731-5p与HIF3A之间的靶向关系,并通过双荧光素酶报告基因实验进行验证。绒毛膜癌细胞中OGFRP1和HIF3A表达增强,而miR-4731-5p表达受到抑制。罗哌卡因处理可抑制绒毛膜癌细胞活力、迁移和侵袭能力。选择用1000μM罗哌卡因处理48小时的绒毛膜癌细胞进行后续实验。OGFRP1升高或miR-4731-5p缺乏可减轻罗哌卡因对绒毛膜癌细胞肿瘤发生的抑制作用。此外,通过StarBase和LncBase预测miR-4731-5p为OGFRP1的潜在靶点,通过StarBase和TargetScan预测HIF3A为miR-4731-5p的潜在靶点。双荧光素酶报告基因实验确定miR-4731-5p是OGFRP1的靶点,HIF3A是miR-4731-5p的靶点。反馈实验表明,miR-4731-5p升高或HIF3A抑制可逆转OGFRP1过表达对罗哌卡因处理的绒毛膜癌细胞恶性行为的促进作用。罗哌卡因通过调节OGFRP1/miR-4731-5p/HIF3A轴抑制绒毛膜癌细胞活力、迁移和侵袭。我们的研究可能为绒毛膜癌的防治提供一种新策略。

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