LINC00839靶向调控miR-124-3p对膀胱癌细胞生物学行为的影响

doi:10.11958/20221488

天津医药 ›› 2023, Vol. 51 ›› Issue (5): 464-467.doi: 10.11958/20221488

LINC00839靶向调控miR-124-3p对膀胱癌细胞生物学行为的影响

吴潇芸¹(), 吴林秀¹, 张丽娣¹, 蓝一笔², 张明津², 易楚繁², 付伟金²^,^△()

1.广西卫生职业技术学院内科护理教研室（邮编530023）
2.广西医科大学第一附属医院泌尿外科

收稿日期:2022-09-15 修回日期:2022-11-04 出版日期:2023-05-15 发布日期:2023-05-05
通讯作者: △E-mail:fuwj66@aliyun.com
作者简介:吴潇芸（1980），女，副教授，主要从事泌尿系肿瘤的基础方面研究。E-mail：1339652193@qq.com
基金资助:
广西卫健委2021年自筹课题(Z20211284);广西卫健委2017年资助课题(S2017034)

The effect of LINC00839 targeting regulation of miR-124-3p on biological behaviors of bladder cancer cells

WU Xiaoyun¹(), WU Linxiu¹, ZHANG Lidi¹, LAN Yibi², ZHANG Mingjin², YI Chufan², FU Weijin²^,^△()

1. Department of Nursing, Guangxi Medical College, Nanning 530023, China
2. Department of Urology, the First Affiliated Hospital of Guangxi Medical University

Received:2022-09-15 Revised:2022-11-04 Published:2023-05-15 Online:2023-05-05
Contact: △E-mail:fuwj66@aliyun.com

吴潇芸, 吴林秀, 张丽娣, 蓝一笔, 张明津, 易楚繁, 付伟金. LINC00839靶向调控miR-124-3p对膀胱癌细胞生物学行为的影响[J]. 天津医药, 2023, 51(5): 464-467.

WU Xiaoyun, WU Linxiu, ZHANG Lidi, LAN Yibi, ZHANG Mingjin, YI Chufan, FU Weijin. The effect of LINC00839 targeting regulation of miR-124-3p on biological behaviors of bladder cancer cells[J]. Tianjin Medical Journal, 2023, 51(5): 464-467.

摘要/Abstract

摘要：

目的探讨LINC00839靶向调控miR-124-3p对膀胱癌细胞生物学行为的影响。方法实时荧光定量PCR（qPCR）检测膀胱癌和癌旁正常组织、膀胱癌细胞株（T24、5637、EJ）中LINC00839表达及抑制LINC00839后对miR-124-3p表达的影响。构建3条小干扰RNA-LINC00839（si-LINC00839-1组、si-LINC00839-2组和si-LINC00839-3组）序列转染膀胱癌细胞。沉默LINC00839表达后，CCK-8检测LINC00839对细胞增殖的影响；Transwell检测LINC00839对细胞侵袭的影响；流式细胞术检测LINC00839对细胞凋亡的影响；双荧光素酶实验检测LINC00839与miR-124-3p的相互作用；免疫蛋白印迹法检测抑制LINC00839后对内皮素受体B（EDNRB）蛋白表达的影响。结果 qPCR检测结果显示，与癌旁正常组织比较，膀胱癌组织中LINC00839表达增高（P<0.05）；LINC00839在膀胱癌细胞5637表达最高。3条siRNA序列中，si-LINC00839-2抑制效率最高。与si-NC组相比，si-LINC00839-2组细胞增殖率下降，侵袭数量减少，细胞凋亡率增加（P<0.05）。双荧光素酶报告检测结果显示，LINC00839与miR-124-3p之间存在互补结合位点。与si-NC组相比，si-LINC00839-2组可上调miR-124-3p表达，抑制EDNRB蛋白表达（P<0.05）。结论 LINC00839可靶向调节miR-124-3p调控膀胱癌细胞增殖、侵袭和凋亡等生物学行为。

关键词: 膀胱肿瘤, 细胞增殖, 细胞凋亡, LINC00839, miR-124-3p

Abstract:

Objective To investigate the effect of LINC00839 targeting regulation of miR-124-3p expression on the biological behavior of bladder cancer cells. Methods Quantitative real-time PCR (qPCR) was used to detect the expression of LINC00839 in bladder cancer and adjacent normal tissue, and bladder cancer cell lines T24, 5637 and EJ. Three small interfering RNA-LINC00839 (si-LINC00839-1, si-LINC00839-2 and si-LINC00839-3) sequences were constructed to transfect bladder cancer cells. After silencing the expression of LINC00839, CCK-8 was used to detect the effect of LINC00839 on cell proliferation. Transwell was used to detect the effect of LINC00839 on cell invasion. The effect of LINC00839 on cell apoptosis was detected by flow cytometry. The interaction between LINC00839 and miR-124-3p was detected by double luciferase assay. Immunoblotting was used to detect the effect of LINC00839 inhibition on endothelial receptor B (EDNRB) protein expression. Results qPCR results showed that the expression of LINC00839 increased significantly in bladder cancer tissue than that in adjacent normal tissue (P<0.05). LINC00839 had the highest expression in bladder cancer 5637 cells. Among three siRNA sequences, si-LINC00839-2 had the highest inhibition efficiency and was used as the experimental group. Compared with the si-NC group, cell proliferation rate, invasion number and apoptosis rate were decreased in the si-LINC00839-2 group (P<0.05). Dual luciferase assay results showed that there were complementary binding sites between LINC00839 and miR-124-3p. Compared with the si-NC group, si-LINC00839-2 group showed the up-regulated expression of miR-124-3p and inhibited expression of EDNRB protein (P<0.05). Conclusion LINC00839 can target miR-124-3p to regulate the biological behaviors of migration, invasion and apoptosis on bladder cancer cells.

Key words: urinary bladder neoplasms, cell proliferation, apoptosis, LINC00839, miR-124-3p

中图分类号:

R737.14

图/表 6

图1 对数生长期的BC细胞（×100）

Fig.1 Bladder cancer cells in logarithmic growth stage (×100)

表1 2组BC细胞不同时间点增殖情况比较（n=3，OD450，$\bar{x}\pm s$）

Tab.1 Comparison of BC cell proliferation at different time points between the two groups

组别	24 h	48 h	72 h	96 h
si-NC组	0.33±0.00	0.62±0.02	1.16±0.03	2.49±0.07
si-LINC00839-2组	0.34±0.00	0.41±0.01	0.67±0.02	1.03±0.04
t	1.957	17.122^＊	23.534^＊	31.981^＊

图2 沉默LINC00839表达对BC5637细胞侵袭的影响（结晶紫染色，×100）

Fig.2 Effects of silencing LINC00839 on the invasion of bladder cancer 5637 cells (crystal violet staining, ×100)

图3 沉默LINC00839表达对BC5637细胞凋亡的影响

Fig.3 Effects of silencing LINC00839 on the apoptosis of bladder cancer 5637 cells

图4 LINC00839与miR-124-3p结合位点

Fig.4 The binding sites of LINC00839 with miR-124-3p

图5 沉默LINC00839对BC5637细胞EDNRB表达的影响

Fig.5 Effects of silencing LINC00839 on the expression of EDNRB protein in bladder cancer 5637 cells

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LINC00839靶向调控miR-124-3p对膀胱癌细胞生物学行为的影响

The effect of LINC00839 targeting regulation of miR-124-3p on biological behaviors of bladder cancer cells

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