miR-625-5p promotes proliferation and invasion of lung adenocarcinoma by targeting PRKACA

胡雅琼; 白　俊; 陈　琳; 陈新璐; 张丽萍; 周丹丹; 王　玉; 尹崇高; 李洪利; 刘雨清

doi:10.19401/j.cnki.1007-3639.2021.06.002

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miR-625-5p promotes proliferation and invasion of lung adenocarcinoma by targeting PRKACA

更新时间：2025-12-31

- miR-625-5p promotes proliferation and invasion of lung adenocarcinoma by targeting PRKACA
- China Oncology Vol. 31, Issue 6, Pages: 447-454(2021)
- 作者机构：
  
  1. 潍坊医学院病理学教研室,山东,潍坊,261053
  2. 潍坊医学院生物科学与技术学院,山东,潍坊,261053
  3. 潍坊医学院护理学院外科护理学教研室,山东,潍坊,261053
  4. 潍坊医学院医学研究实验中心,山东,潍坊,261053
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2021.06.002
  CLC：
- Published Online：09 July 2021，
  
  Published：09 July 2021
- 稿件说明：
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胡雅琼, 白　俊, 陈　琳, 陈新璐, 张丽萍, 周丹丹, 王　玉, 尹崇高, 李洪利, 刘雨清 . miR-625-5p通过靶向调控PRKACA促进肺腺癌细胞的增殖和侵袭[J]. 中国癌症杂志, 2021, 31(6): 447-454.

胡雅琼, 白　俊, 陈　琳, et al. miR-625-5p promotes proliferation and invasion of lung adenocarcinoma by targeting PRKACA[J]. China Oncology, 2021, 31(6): 447-454.
胡雅琼, 白　俊, 陈　琳, 陈新璐, 张丽萍, 周丹丹, 王　玉, 尹崇高, 李洪利, 刘雨清 . miR-625-5p通过靶向调控PRKACA促进肺腺癌细胞的增殖和侵袭[J]. 中国癌症杂志, 2021, 31(6): 447-454. DOI： 10.19401/j.cnki.1007-3639.2021.06.002.

胡雅琼, 白　俊, 陈　琳, et al. miR-625-5p promotes proliferation and invasion of lung adenocarcinoma by targeting PRKACA[J]. China Oncology, 2021, 31(6): 447-454. DOI： 10.19401/j.cnki.1007-3639.2021.06.002.

摘要

背景与目的：肺腺癌是非小细胞肺癌的一种亚型，虽在诊断和治疗方面已经取得了很大进展，但晚期肺腺癌临床预后和总生存仍较差。近年来多项研究表明，miRNA在多种癌症中发挥作用，并在细胞增殖、转移、炎症等生物学过程中发挥重要作用。探究miR-625-5p对肺腺癌细胞增殖和侵袭能力的影响及分子机制，旨在为后续肺腺癌的诊断和治疗提供新的思路。方法：利用GEO数据库查找肺腺癌组织中差异表达的miRNA。采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）检测miR-625-5p在各肺腺癌细胞系中的表达情况，采用EdU细胞增殖实验和transwell侵袭实验研究miR-625-5p对肺腺癌细胞增殖和侵袭能力的影响；通过生物信息学预测miR-625-5p靶向结合的关键基因；采用蛋白质印迹法（Western blot）检测对蛋白激酶cAMP激活催化亚基α（protein kinase cAMP-activated catalytic subunit alpha，PRKACA）在肺腺癌细胞中的表达情况进行验证，采用双荧光素酶报告基因实验和Western blot分析miR-625-5p与PRKACA之间的靶向关系。Western blot检测共转染敲低PRKACA质粒和敲低miR-625-5p质粒后各组细胞PRKACA的表达情况。采用EdU细胞增殖实验和transwell侵袭实验检测共转染以后各组肺腺癌细胞增殖和侵袭能力的改变。结果：miR-625-5p在肺腺癌组织（P0.000 1）和各组肺腺癌细胞（P0.000 1）中表达上调。EdU细胞增殖实验和transwell侵袭实验结果显示，miR-625-5p能够促进肺腺癌细胞的增殖（P=0.002 3）和侵袭（P=0.000 3）能力。双荧光素酶报告基因实验结果显示，miR-625-5p能与PRKACA靶向结合（P=0.000 8）。在肺腺癌细胞中，miR-625-5p与PRKACA的表达呈负相关（P0.000 1）。miR-625-5p下调能够逆转敲低PRKACA对A549细胞增殖（P=0.011 9）和侵袭（P=0.001 5）能力的促进作用。结论：miR-625-5p在肺腺癌组织和细胞中表达上调，并通过负向调控PRKACA促进肺腺癌细胞增殖和侵袭。

Abstract

Background and purpose: Lung adenocarcinoma is a subtype of non-small cell lung cancer. Although much progress has been made in the diagnosis and treatment of lung adenocarcinoma

the clinical prognosis and overall survival of advanced lung adenocarcinoma are still poor. In recent years

a number of studies have shown that miRNA can play a role in a variety of cancers

and play an important role in cell proliferation

metastasis

inflammation and other biological processes. This study aimed to explore the effect of miR-625-5p on the proliferation and invasion ability of lung adenocarcinoma cells and its molecular mechanism

so as to provide a new idea for the diagnosis and treatment of lung cancer. Methods: GEO database was used to search for differentially expressed miRNA in lung adenocarcinoma. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression of miR-625-5p in various lung adenocarcinoma cell lines. The effects of miR-625-5p on the proliferation and invasion of lung adenocarcinoma cells were investigated by EdU cell proliferation assay and transwell invasion assay. Key genes of miR-625-5p targeted binding were predicted by bioinformatics. Western blot experiment validated the expression of protein kinase cAMP-activated catalytic subunit alpha (PRKACA) in lung adenocarcinoma cells. The targeted relationship between miR-625-5p and PRKACA was analyzed by double luciferase assay and Western blot. Western blot assay was used to detect the expression of PRKACA after co-transfection of si-PRKACA and si-miR-625-5p in each group. The effect of miR-625-5p on the proliferation and invasion ability of lung adenocarcinoma cells by targeting PRKACA was observed by EdU cell proliferation assay and transwell invasion assay. Results: The expression of miR-625-5p was up-regulated in lung adenocarcinoma tissues (P0.000 1) and cells (P0.000 1). The results of EdU cell proliferation assay and transwell invasion assay showed that miR-625-5p promoted the proliferation (P=0.002 3) and invasion (P=0.000 3) of lung adenocarcinoma cells. Double luciferase assay showed that miR-625-5p could target and bind to PRKACA (P=0.000 8). In lung adenocarcinoma cells

miR-625-5p was negatively correlated with PRKACA expression (P0.000 1). Down-regulation of miR-625-5p reversed the promotion of the proliferation (P=0.011 9) and invasion (P=0.001 5) ability of A549 cells by knockout of PRKACA. Conclusion: MiR-625-5p is up-regulated in lung adenocarcinoma and promotes proliferation and invasion of lung adenocarcinoma tissues and cells by negatively regulating PRKACA.

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