LncRNA SNHG5/miR-26a-5p/MTDH信号轴促进结直肠癌转移的机制研究

冶俊玲; 郑小影; 郭新建; 陈瑞慧; 杨柳; 苟笑丹; 蒋汉梅

doi:10.19401/j.cnki.1007-3639.2023.07.005

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LncRNA SNHG5/miR-26a-5p/MTDH信号轴促进结直肠癌转移的机制研究

论著 | 更新时间：2025-12-31

- LncRNA SNHG5/miR-26a-5p/MTDH信号轴促进结直肠癌转移的机制研究
- A study on mechanism of lncRNA-mediated SNHG5/miR-26a-5p/MTDH signal axis promoting metastasis of colorectal cancer
- 中国癌症杂志 2023年33卷第7期页码：673-685
- 作者机构：
  
  1. 青海大学附属医院病理科，青海西宁 810001
  2. 青海大学附属医院消化内科，青海西宁 810001
- 作者简介：
  
  冶俊玲（ORCID: 0000-0002-4899-4353），本科，副主任医师。
- 基金信息：
  
  青海省卫生健康委员会一般指导性课题(2020-wjzdx-46)
- DOI：10.19401/j.cnki.1007-3639.2023.07.005
  中图分类号： R735.3
- 收稿：2022-09-22，
  
  修回：2023-01-03，
  
  纸质出版：2023-07-30
- 稿件说明：
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冶俊玲, 郑小影, 郭新建, 等. LncRNA SNHG5/miR-26a-5p/MTDH信号轴促进结直肠癌转移的机制研究[J]. 中国癌症杂志, 2023,33(7):673-685.

Junling YE, Xiaoying ZHENG, Xinjian GUO, et al. A study on mechanism of lncRNA-mediated SNHG5/miR-26a-5p/MTDH signal axis promoting metastasis of colorectal cancer[J]. China Oncology, 2023, 33(7): 673-685.
冶俊玲, 郑小影, 郭新建, 等. LncRNA SNHG5/miR-26a-5p/MTDH信号轴促进结直肠癌转移的机制研究[J]. 中国癌症杂志, 2023,33(7):673-685. DOI： 10.19401/j.cnki.1007-3639.2023.07.005.

Junling YE, Xiaoying ZHENG, Xinjian GUO, et al. A study on mechanism of lncRNA-mediated SNHG5/miR-26a-5p/MTDH signal axis promoting metastasis of colorectal cancer[J]. China Oncology, 2023, 33(7): 673-685. DOI： 10.19401/j.cnki.1007-3639.2023.07.005.

摘要

背景与目的：

长链非编码RNA小核仁RNA宿主基因5（long non-coding RNA small nucleolar RNA host gene 5，lncRNA SNHG5）在多种癌症中发挥促癌作用，但其对结直肠癌（colorectal cancer，CRC）的影响和调节机制尚不清楚。本研究旨在探究lncRNA SNHG5/miR-26a-5p/异粘蛋白（metadherin，MTDH）信号轴促进CRC转移的机制。

方法：

分析癌症基因组图谱（The Cancer Genome Atlas，TCGA）数据库中的数据，探索CRC中异常表达的lncRNA并进行生存分析。收集2020年10月—2021年10月经手术切除的100例CRC、癌旁组织样本及患者的完整临床资料，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lncRNA SNHG5、miR-26a-5p表达水平，采用免疫组织化学法检测MTDH的表达水平，分析lncRNA SNHG5在CRC中的相对表达水平与临床病理学特征及生存期的关系。通过细胞计数试剂盒-8（cell counting kit-8，CCK-8）检测、克隆形成、划痕、transwell实验及体内异种移植实验检测lncRNA SNHG5对CRC细胞增殖、迁移及侵袭的影响，通过蛋白质印迹法（Western blot）和免疫组织化学检测CRC细胞转移、上皮-间充质转化相关分子表达水平与lncRNA SNHG5表达水平的关系。通过RNA下拉实验、双荧光素酶报告基因检测、RNA免疫共沉淀实验探究SNHG5与miR-26a-5p、MTDH与miR-26a-5p在物理上的相互作用，通过CCK-8、EdU、transwell实验验证三者在功能上的相互关系，采用Western blot检测分析SNHG5、miR-26a-5p、MTDH表达对迁移、侵袭相关分子的影响。

结果：

TCGA数据库分析结果显示，lncRNA SNHG5在CRC中显著上调。RTFQ-PCR和免疫组织化学检测结果显示，CRC组织中lncRNA SNHG5、MTDH水平显著上调（

0.05），miR-26a-5p水平下降（

0.05），SNHG5高表达的样本中MTDH水平也较高。浆膜及浆膜外浸润、远处转移、淋巴结转移、TNM Ⅲ期的CRC组织lncRNA SNHG5表达量高于浆膜下浸润、无远处转移、无淋巴结转移、TNM Ⅰ~Ⅱ期的组织，差异有统计学意义（

0.05）。生存分析结果显示，lncRNA SNHG5高表达与总生存率显著相关（

0.05）。过表达lncRNA SNHG5能够提升CRC细胞增殖、克隆形成、迁移及侵袭能力，促进移植瘤生长和肺转移，提高Ki-67增殖指数和波形蛋白（vimentin）的相对表达水平（

0.05），降低E-钙粘蛋白（E-cadherin）的相对表达水平（

0.05），而抑制lncRNA SNHG5表达后CRC细胞的发展受到抑制。RNA下拉实验、双荧光素酶报告基因检测、RNA免疫共沉淀实验证实SNHG5与miR-26a-5p、MTDH及miR-26a-5p存在物理上的相互作用，在lncRNA SNHG5过表达细胞中上调miR-26a-5p或下调MTDH表达可部分逆转lncRNA SNHG5对CRC细胞增殖、迁移、侵袭及相关分子表达水平的影响。

结论：

LncRNA SNHG5在CRC组织和细

胞中表达上调，其高表达与肿瘤进展及生存不良有关，可作为miR-26a-5p的分子海绵调节MTDH表达促进SW620细胞增殖和转移。

Abstract

Background and purpose:

Long non-coding RNA small nucleolar RNA host gene 5 (lncRNA SNHG5) plays a cancer-promoting role in many cancers

however its effect on colorectal cancer (CRC) and its regulatory mechanism are not clear. This study aimed to explore the mechanism of lncRNA SNHG5/miR-26a-5p/metadherin (MTDH) signal axis promoting metastasis of CRC.

Methods:

The data of The Cancer Genome Atlas (TCGA) database was analyzed

the abnormal expression of lncRNA in CRC was explored and analyzed the survival. Samples of CRC

paracancerous tissues and complete clinical data of patients who underwent surgical resection from October 2020 to October 2021 were collected. The expression levels of SNHG5 and miR-26a-5p in lncRNA were detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR)

and the expression level of MTDH was detected by immunohistochemistry. The relationship between the relative expression level of lncRNA SNHG5 in CRC and clinicopathological features and survival time was analyzed. The effects of lncRNA SNHG5 on the proliferation

migration and invasion of CRC cells were detected by cell counting kit-8 (CCK-8)

clone formation

scratching assays

transwell test and in vivo xenotransplantation. The relationship between CRC cell metastasis

the expression level of epithelial-mesenchymal transition related molecules and lncRNA SNHG5 expression level by Western blot and immunohistochemical detection were explored. The physical interaction between SNHG5 and miR-26a-5p

MTDH and miR-26a-5p was studied by RNA pull-down test

double luciferase reporter gene detection and RNA co-immunoprecipitation. The functional relationship among the three was verified by CCK-8

EdU and transwell experiments. The effect of SNHG5

miR-26a-5p and MTDH expression on migration and invasion related molecules was analyzed by West

ern blot.

Results:

The results of TCGA database analysis showed that lncRNA SNHG5 was significantly upregulated in CRC. The results of RTFQ-PCR and immunohistochemistry showed that the levels of lncRNA SNHG5 and MTDH in CRC tissues were significantly upregulated (

0.05)

the level of miR-26a-5p was decreased (

0.05)

and the level of MTDH in samples with high expression of SNHG5 was also increased. The expression of lncRNA SNHG5 in CRC tissues with serosa and extraserosal invasion

distant metastasis

lymph node metastasis and TNM stage Ⅲ was significantly higher compared with subserosal invasion

no distant metastasis and lymph node metastasis and TNM stage Ⅰ-Ⅱ (

0.05). The results of survival analysis showed that the high expression of lncRNA SNHG5 was significantly correlated with overall survival rate (

0.05). Overexpression of lncRNA SNHG5 could enhance the proliferation

clone formation

migration and invasion of CRC cells

promote the growth and lung metastasis of transplanted tumor

increase the relative expression level of Ki-67 proliferation index and vimentin (

0.05)

and decrease the relative expression level of E-cadherin (

0.05). However

the development of CRC cells was inhibited after inhibition of lncRNA SNHG5 expression. RNA pull-down test

double luciferase reporter gene detection and RNA co-immunoprecipitation confirmed the physical interaction between SNHG5 and miR-26a-5p

MTDH and miR-26a-5p. Upregulation of miR-26a-5p or downregulation of MTDH expression in lncRNA SNHG5 overexpressed cells partially reversed the effects of lncRNA SNHG5 on proliferation

migration

invasion and expression of related molecules in CRC cells.

Conclusion:

LncRNA SNHG5 is upregulated in CRC tissues and cells

and its high expression is related to tumor progression and poor survival. It can be used as a molecular sponge of miR-26a-5p to regulate the expression of M

TDH to promote the proliferation and metastasis of SW620 cells.

关键词

Keywords

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