miR-196b靶向ERG促进肺腺癌的增殖和迁移

刘邦卿; 李剑锋; 刘晓辉; 张劲男; 梁金屏

doi:10.12259/j.issn.2095-610X.S20231023

miR-196b靶向ERG促进肺腺癌的增殖和迁移

doi: 10.12259/j.issn.2095-610X.S20231023

1.
唐山市人民医院胸外二科
2.
头颈外科，河北唐山　063000

基金项目: 河北省医学科学研究课题基金资助项目（20231805）

详细信息

作者简介:
刘邦卿（1988～），男，山东巨野人，医学硕士，主治医师，主要从事肺癌基础与临床研究工作

中图分类号: R734.2
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出版历程
- 收稿日期: 2023-06-26
- 刊出日期: 2023-10-25

MiR-196b Promotes Proliferation and Migration of Lung Adenocarcinoma by Targeting ERG

1.
The 2nd Dept. of Thoracic Surgery
2.
Dept. of Head and Neck Surgery，Tangshan People’s Hospital，Tangshan Hebei 063000，China

摘要

摘要: 目的探究miR-196b影响肺腺癌（lung adenocarcinoma，LUAD）进展的机制。方法通过TCGA数据库分析miR-196b在LUAD组织中的表达水平，利用TargetScan预测其下游靶基因并用双荧光素酶实验进行验证，qRT-PCR检测miR-196b和ETS相关基因（ETS-related gene，ERG）在LUAD细胞系中的表达情况，MTT、划痕愈合和Transwell实验分别检测不同处理后LUAD细胞的增殖、迁移和侵袭能力的变化情况。结果 miR-196b在LUAD中高表达（P = 3.50e-17）并靶向负调控ERG，过表达miR-196b能够促进LUAD细胞的增殖、迁移和侵袭（P < 0.05），回复实验证实miR-196b/ERG调控轴能够影响LUAD细胞增殖、迁移和侵袭（ P < 0.05）。结论 miR-196b靶向ERG促进LUAD进展的分子机制，对开发LUAD新的临床治疗方法具有潜在意义。
- miR-196b /
- ERG /
- 肺腺癌 /
- 增殖 /
- 迁移
Abstract: Objective To explore the mechanisms of miR-196b affecting the progression of lung adenocarcinoma （LUAD）. Methods The expression level of miR-196b in LUAD tissues was analyzed through TCGA database. TargetScan was used to predict its downstream target genes, and dual-luciferase assay was performed to validate the predictions. qRT-PCR was used to detect the expression of miR-196b and ETS-related gene （ERG） in LUAD cell lines. MTT, scratch healing, and Transwell assays were used to respectively assess the changes in proliferation, migration, and invasion abilities of LUAD cells after different treatments. Results miR-196b was highly expressed in LUAD （P = 3.50e-17） and negatively regulated ERG. Overexpression of miR-196b promoted proliferation, migration, and invasion of LUAD cells （P < 0.05）. Rescue experiments confirmed that miR-196b/ERG regulatory axis can affect the proliferation, migration, and invasion of LUAD cells （ P < 0.05）. Conclusion The molecular mechanism of miR-196b targeting ERG in promoting the progression of LUAD has potential significance for the development of new clinical treatment methods for LUAD.
- miR-196b /
- ERG /
- Lung adenocarcinoma /
- Proliferation /
- Migration

HTML全文

图 1 miR-196b和ERG在肺腺癌中的表达情况

A：通过TCGA数据库预测miR-196b在正常组织和肺腺癌组织中的表达，绿框图表示正常样本（n = 46），红框图表示肿瘤样本（n = 521）；B：肺腺癌患者miR-196b表达水平与总生存率的K-M分析，红色表示高表达组，蓝色表示低表达组；C：通过TCGA数据库预测正常组织和肺腺癌组织中ERG的表达，绿框图表示正常样本（n = 59），红框图表示肿瘤样本（n = 535）。

Figure 1. Expression of miR-196b and ERG in lung adenocarcinoma

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图 2 miR-196b在肺腺癌细胞中的表达情况

与BEAS-2B组比较，^*P < 0.05。

Figure 2. Expression of miR-196b in lung adenocarcinoma cells

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图 3 miR-196b可影响肺腺癌细胞的增殖、迁移和侵袭

A：qRT-PCR检测过表达或敲低miR-196b后细胞的转染效率；B：MTT法检测过表达或敲低miR-196b后细胞的增殖能力；C：划痕愈合实验检测过表达或敲低miR-196b后细胞的迁移能力；D：Transwell实验检测过表达或敲低miR-196b后细胞的迁移和侵袭能力（100×）。与NC组比较，^*P < 0.05。

Figure 3. miR-196b can affect the proliferation，migration and invasion of lung adenocarcinoma cells

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图 4 miR-196b的靶点检测及不同处理组细胞中ERG 的表达情况

A：通过TargetScan数据库预测miR-196b和ERG的靶向结合位点；B：双荧光素酶法检测miR-196b与ERG的靶向结合关系；C：采用qRT-PCR和Western blot检测各细胞系中ERG mRNA和蛋白的表达；D：采用qRT-PCR和Western blot检测不同处理组ERG mRNA和蛋白的表达。与NC组或BEAS-2B组做比较，^*P < 0.05。

Figure 4. Target detection of miR-196b and expression of ERG in cells of different treatment groups

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图 5 miR-196b能靶向下调ERG促进肺腺癌细胞的增殖、迁移和侵袭能力

A：采用qRT-PCR和Western blot检测不同处理组大鼠ERG mRNA和蛋白的表达；B：MTT实验检测不同处理组的细胞增殖能力；C：划痕愈合实验检测不同处理组细胞迁移能力；D：Transwell实验检测不同处理组细胞的迁移和侵袭能力（100×）。NC-mimic、NC-inhibitor、oe-NC和si-NC分别表示miR-196b过表达、miR-196b沉默、ERG过表达和ERG沉默对照组，miR-mimic和miR-inhibitor分别表示miR-196b过表达和沉默组，oe-ERG和si-ERG分别表示ERG过表达组和沉默组；miR-mimic/inhibitor + oe/si-NC与NC组比较，miR-mimic/inhibitor + oe/si-NC组与miR-mimic/inhibitor + oe/si-ERG组比较，^*P < 0.05，。

Figure 5. miR-196b can target down regulate ERG to promote the proliferation，migration and invasion of lung adenocarcinoma cells

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表 1 研究使用的细胞系及培养基

Table 1. Cell lines and media used in the study

细胞系	编号	培养基
BEAS-2B	BNCC338205	DMEM-H
PC-9	BNCC340767	DMEM-H
HCC-78	BNCC338064	RPMI-1640
H1395	BNCC255519
Calu-3	BNCC338514

下载: 导出CSV

表 2 引物序列表

Table 2. Primer Sequence Table

基因	引物序列（5′→ 3′）
miR-196b	Forward primer: GTACCACTTTATCCCGTTCACCA Reverse primer:ATCTCGAGGCAGGGAGAGAGGAATAA
U6	Forward primer: GCCCATCTTGACCCGAAT
U6	Reverse primer: AACGCTTCACGAATTTGCGT
ERG	Forward primer:GAGTGGGCGGTGAAAGAATA
ERG	Reverse primer: GGAGATGTGAGAGAAGAGTG
GAPDH	Forward primer:CTCCTCCTGTTCGACAGTCAGC
GAPDH	Reverse primer:CCCAATACGACCAAATCCGTT

下载: 导出CSV

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