M6A甲基化调控因子对结直肠癌预后及细胞生物学行为的影响

刘宁; 江帆; 陈之巨; 王葆春; 吕云福

doi:10.16016/j.2097-0927.202111061

您当前的位置：

首页 >

文章列表页 >

M6A甲基化调控因子对结直肠癌预后及细胞生物学行为的影响

基础医学 | 更新时间：2026-01-09

- M6A甲基化调控因子对结直肠癌预后及细胞生物学行为的影响
- Effects of N6-methyladenosine methylation regulators on prognosis and cell biological behaviors of colorectal cancer
- 陆军军医大学学报 2022年44卷第11期页码：1126-1135
- 作者机构：
  
  570311 海口, 海南省人民医院胃肠外一科
- 作者简介：
  
  [刘宁，E-mail: drliuning@hainmc.edu.cnLIU Ning, E-mail: drliuning@hainmc.edu.cn
- 基金信息：
  
  海南省自然科学基金青年基金(819QN353);海南省重点研发项目(ZDYF2022SHFZ116);海南省人民医院院级项目(2021MSXM07)
- DOI：10.16016/j.2097-0927.202111061
  中图分类号： R329.28;R730.23;R735.2
- 收稿：2021-11-05，
  
  修回：2022-01-03，
  
  纸质出版：2022-06-15
- 稿件说明：
移动端阅览
刘宁, 江帆, 陈之巨, 等. M6A甲基化调控因子对结直肠癌预后及细胞生物学行为的影响[J]. 陆军军医大学学报, 2022,44(11):1126-1135.

Ning LIU, Fan JIANG, Zhiju CHEN, et al. Effects of N6-methyladenosine methylation regulators on prognosis and cell biological behaviors of colorectal cancer[J]. Journal of Army Medical University, 2022, 44(11): 1126-1135.
刘宁, 江帆, 陈之巨, 等. M6A甲基化调控因子对结直肠癌预后及细胞生物学行为的影响[J]. 陆军军医大学学报, 2022,44(11):1126-1135. DOI： 10.16016/j.2097-0927.202111061.

Ning LIU, Fan JIANG, Zhiju CHEN, et al. Effects of N6-methyladenosine methylation regulators on prognosis and cell biological behaviors of colorectal cancer[J]. Journal of Army Medical University, 2022, 44(11): 1126-1135. DOI： 10.16016/j.2097-0927.202111061.

摘要

目的

探讨N6-甲基腺苷(N6-methyladenosine，m6A)相关基因对结直肠癌预后及细胞生物学行为的影响。

方法

从肿瘤基因组图谱(the cancer genome atlas，TCGA)下载长链非编码RNA(long noncoding RNA

lncRNA)和21个M6A相关基因的表达谱数据，R软件分析差异基因；聚类分析和主成分分析(principal component analysis，PCA)分析M6A相关基因；单因素Cox回归分析筛选预后相关M6A基因，LASSO算法建立风险预测模型；筛选结直肠癌相关差异lncRNA(differentially expressed lncRNA，DElncRNA)，与M6A相关基因建立共表达网络。使用实时荧光定量PCR(real time fluorescence quantitative PCR，RT-qPCR)检测M6A reader脆性X智力低下1(fragile X mental retardation 1，FMR1)在结直肠癌组织中的表达，在结直肠癌细胞中敲除FMR1基因，通过CCK-8、划痕实验、Transwell和细胞凋亡实验检测细胞的增殖、迁移、侵袭和凋亡情况。

结果

21个M6A相关基因中有15个基因在结直肠癌中差异表达。聚类分析显示，结直肠癌肿瘤分成2种亚型；从建立的风险模型得出烷基化修复同源蛋白5(a-ketoglutarate-dependent dioxygenase alk B homolog 5，ALKBH5)、YTH结构域蛋白2(YTH domaincontaining protein 2，YTHDC2)和FMR1可能是独立的预后因子。随后，筛选出1 784个DElncRNA，构建包含3个预后因子和18个DElncRNAs的共表达网络。网络图中发现FMR1基因占主导地位，RT-qPCR实验发现FMR1在结直肠癌组织中上调；FMR1基因的敲除能抑制结直肠癌HT-29细胞的增殖，迁移和侵袭能力，促进结直肠癌细胞的凋亡。

结论

M6A甲基化调控因子ALKBH5、YTHDC2和FMR1可能是结直肠癌中的独立预后因子，FMR1基因的敲除能抑制结直肠癌细胞的增殖、迁移、侵袭，促进结直肠癌细胞的凋亡。

Abstract

Objective

To investigate the effects of N6-methyladenosine (M6A) related genes on the prognosis and cell biological behaviors of colorectal cancer.

Methods

The long non-coding RNA (lncRNA) for colorectal cancer and the expression profile data of 21 M6A-related genes were downloaded from The Cancer Genome Atlas (TCGA)

and the differential genes were analyzed with R software. Cluster analysis and principal component analysis (PCA) were further conducted

and prognostic related M6A genes were screened by univariate Cox regression analysis. The risk prediction model was then established with LASSO algorithm. In addition

differentially expressed lncRNA associated with colorectal cancer (DElncRNA) were screened out

and a co-expression network with M6A-related genes was subsequently established. Moreover

RT-qPCR was performed to determine the gene expression of fragile X mental retardation 1 (FMR1) in colorectal cancer tissues. After FMR1 gene was knocked out in colorectal cancer cells

the cell proliferation

migration

invasion and apoptosis were detected by CCK8 assay

cell scratch test

Transwell assay and flow cytometry

respectively.

Results

Fifteen of the 21 M6A-related genes were differentially expressed in colorectal cancer

and cluster analysis indicated 2 subtypes of colorectal cancer tumors. According to the risk model

ALKBH5 (a-ketoglutarate-dependent dioxygenase alk B homolog 5)

YTHDC2 (YTH domain-containing protein 2) and FMR1 may be the independent prognostic factors. With a total of 1 784 DElncRNAs screened out

a co-expression network was successfully constructed containing the 3 prognostic factors and 18 DElncRNAs. FMR1 was found to be the hub gene in the network diagram. RT-qPCR showed that FMR1 was up-regulated in colorectal cancer tissues. Knockout of FMR1 gene inhibited the proliferation

migration and invasion

and promoted cell apoptosis in colorectal cancer HT-29 cells.

Conclusion

M6A methylation regulators ALKBH5

YTHDC2 and FMR1 may be the independent prognostic factors in colorectal cancer. FMR1 knockout can inhibit the proliferation

migration and invasion

and promote the apoptosis of colorectal cancer cells.

关键词

Keywords

references

FERLAY J, STELIAROVA-FOUCHER E, LORTET-TIEULENT J, et al . Cancer incidence and mortality patterns in Europe: Estimates for 40 countries in 2012[J ] . Eur J Cancer, 2013, 49(6): 1374-1403. DOI:10.1016/j.ejca.2012.12.027.

FERLAY J, SOERJOMATARAM I, DIKSHIT R, et al . Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012[J ] . Int J Cancer, 2015, 136(5): E359-E386. DOI:10.1002/ijc.29210.

王唯全, 李萍, 王楚盈, 等. 结直肠癌的病因病机与药物治疗的研究进展[J]. 长春中医药大学学报, 2020, 36(1): 194-197. DOI:10.13463/j.cnki.cczyy.2020.01.053.

WANG W Q, LI P, WANG C Y, et al . Advances in etiology, pathogenesis and drug therapy of colorectal cancer[J ] . J Changchun Univ Chin Med, 2020, 36(1): 194-197. DOI:10.13463/j.cnki.cczyy.2020.01.053.

INADOMI J M. Screening for colorectal neoplasia[J]. N Engl J Med, 2017, 376(2): 149-156. DOI:10.1056/NEJMcp1512286.

HUANG H, WENG H, CHEN J. The biogenesis and precise control of RNA M6A methylation[J]. Trends Genet, 2020, 36(1): 44-52. DOI:10.1016/j.tig.2019.10.011.

HUANG H, WENG H, SUN W, et al . Recognition of RNA N 6 -methyladenosine by IGF2BP proteins enhances mRNA stability and translation[J ] . Nat Cell Biol, 2018, 20(3): 285-295. DOI:10.1038/s41556-018-0045-z..

ROUNDTREE I A, EVANS M E, PAN T, et al . Dynamic RNA modifications in gene expression regulation[J ] . Cell, 2017, 169(7): 1187-1200. DOI:10.1016/j.cell.2017.05.045.

SUI X, HU Y, REN C, et al . METTL3-mediated M6A is required for murine oocyte maturation and maternal-to-zygotic transition[J ] . Cell Cycle, 2020, 19(4): 391-404. DOI:10.1080/15384101.2019.1711324.

LIU L, WANG J, SUN G, et al . M6A mRNA methylation regulates CTNNB 1 to promote the proliferation of hepatoblastoma[J ] . Mol Cancer, 2019, 18(1): 188. DOI:10.1186/s12943-019-1119-7..

LIN S, CHOE J, DU P, et al . The m(6)A methyltransferase METTL3 promotes translation in human cancer cells[J ] . Mol Cell, 2016, 62(3): 335-345. DOI:10.1016/j.molcel.2016.03.021.

LIU J, ECKERT M A, HARADA B T, et al . M6A mRNA methylation regulates AKT activity to promote the proliferation and tumorigenicity of endometrial cancer[J ] . Nat Cell Biol, 2018, 20(9): 1074-1083. DOI:10.1038/s41556-018-0174-4.

CHENG X, LI M, RAO X, et al . KIAA1429 regulates the migration and invasion of hepatocellular carcinoma by altering M6A modification of ID2 mRNA[J ] . Onco Targets Ther, 2019, 12: 3421-3428. DOI:10.2147/ott.s180954.

WU Y, YANG X, CHEN Z, et al . M6A-induced lncRNA RP11 triggers the dissemination of colorectal cancer cells via upregulation of Zeb1[J ] . Mol Cancer, 2019, 18(1): 87. DOI:10.1186/s12943-019-1014-2..

SIEGEL R L, MILLER K D, JEMAL A. Cancer statistics, 2018[J]. CA Cancer J Clin, 2018, 68(1): 7-30. DOI:10.3322/caac.21442.

UNSELD M, DRIMMEL M, SIEBENHÜNER A, et al . Optimizing treatment sequence for late-line metastatic colorectal cancer patients using trifluridine/tipiracil and regorafenib[J ] . Clin Colorectal Cancer, 2018, 17(4): 274-279. DOI:10.1016/j.clcc.2018.05.012.

ZHANG S, ZHAO B S, ZHOU A, et al . M6A demethylase ALKBH 5 maintains tumorigenicity of glioblastoma stem-like cells by sustaining FOXM1 expression and cell proliferation program[J ] . Cancer Cell, 2017, 31(4): 591-606. e6. DOI:10.1016/j.ccell.2017.02.013..

CAI X, WANG X, CAO C, et al . HBXIP-elevated methyltransferase METTL3 promotes the progression of breast cancer via inhibiting tumor suppressor let-7g[J ] . Cancer Lett, 2018, 415: 11-19. DOI:10.1016/j.canlet.2017.11.018.

GUO T, LIU D F, PENG S H, et al . ALKBH 5 promotes colon cancer progression by decreasing methylation of the lncRNA NEAT1[J ] . Am J Transl Res, 2020, 12(8): 4542-4549..

YUAN Y, YAN G, HE M, et al . ALKBH 5 suppresses tumor progression via an M6A-dependent epigenetic silencing of pre-miR-181b-1 /YAP signaling axis in osteosarcoma[J ] . Cell Death Dis, 2021, 12(1): 60. DOI:10.1038/s41419-020-03315-x..

MA L, CHEN T, ZHANG X, et al . The M6A reader YTHDC2 inhibits lung adenocarcinoma tumorigenesis by suppressingSLC7A11-dependent antioxidant function[J ] . Redox Biol, 2021, 38: 101801. DOI:10.1016/j.redox.2020.101801.

SUN S, HAN Q, LIANG M, et al . Downregulation of m 6 A reader YTHDC2 promotes tumor progression and predicts poor prognosis in non-small cell lung cancer[J ] . Thorac Cancer, 2020, 11(11): 3269-3279. DOI:10.1111/1759-7714.13667..

EDUPUGANTI R R, GEIGER S, LINDEBOOM R G H, et al . N 6 -methyladenosine (M6A) recruits and repels proteins to regulate mRNA homeostasis[J ] . Nat Struct Mol Biol, 2017, 24(10): 870-878. DOI:10.1038/nsmb.3462..

NI W, YAO S, ZHOU Y X, et al . Long noncoding RNA GAS5 inhibits progression of colorectal cancer by interacting with and triggering YAP phosphorylation and degradation and is negatively regulated by the M6A reader YTHDF 3 [J ] . Mol Cancer, 2019, 18(1): 143. DOI:10.1186/s12943-019-1079-y..

LI T, HU P S, ZUO Z, et al . METTL3 facilitates tumor progression via an M6A-IGF 2 BP 2 -dependent mechanism in colorectal carcinoma[J ] . Mol Cancer, 2019, 18(1): 112. DOI:10.1186/s12943-019-1038-7..

ZHENG Y, NIE P, XU S. Long noncoding RNA CASC21 exerts an oncogenic role in colorectal cancer through regulating miR-7-5p/YAP1 axis[J]. Biomed Pharmacother, 2020, 121: 109628. DOI:10.1016/j.biopha.2019.109628.

WANG L, DUAN W, YAN S, et al . Circulating long non-coding RNA colon cancer-associated transcript 2 protected by exosome as a potential biomarker for colorectal cancer[J ] . Biomed Pharmacother, 2019, 113: 108758. DOI:10.1016/j.biopha.2019.108758.

XU J, MENG Q T, LI X B, et al . Long noncoding RNA MIR17HG promotes colorectal cancer progression via miR-17-5p[J ] . Cancer Res, 2019, 79(19): 4882-4895. DOI:10.1158/0008-5472.CAN-18-3880.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

基于中国人群的雌激素受体表达新分层（低/中/高）可更精准地预测HR⁺/HER2^-早期乳腺癌患者的预后

转录因子TGIF1高表达胃癌的免疫抑制性微环境特征及临床意义分析

核仁磷酸蛋白1突变的分子异质性对急性髓系白血病预后的影响

5因子改良衰弱指数联合控制营养状态评分与不可切除肝癌患者介入治疗后总生存期的相关性分析

全身炎症反应指数联合临床病理参数预测宫颈癌复发的模型建立与验证