长链非编码RNA LINC00601调控肝细胞癌奥沙利铂耐药的分子机制研究

胡可舒; 刘文凤; 张锋; 权冰; 殷欣

doi:10.19401/j.cnki.1007-3639.2023.01.003

您当前的位置：

首页 >

文章列表页 >

长链非编码RNA LINC00601调控肝细胞癌奥沙利铂耐药的分子机制研究

论著 | 更新时间：2025-12-31

- 长链非编码RNA LINC00601调控肝细胞癌奥沙利铂耐药的分子机制研究
- Mechanism of LINC00601 in regulating sensitivity of hepatocellular carcinoma cells to oxaliplatin chemotherapy
- 中国癌症杂志 2023年33卷第1期页码：25-35
- 作者机构：
  
  1. 复旦大学附属中山医院肝肿瘤内科，上海 200032
  2. 复旦大学附属中山医院肝癌研究所，上海 200032
- 作者简介：
  
  [ "胡可舒（ORCID: 0000-0003-4576-3688），硕士。" ]
  殷欣（ORCID: 0000-0002-9892-7845），博士，副主任医师。
- 基金信息：
  
  国家自然科学基金(81672331)
- DOI：10.19401/j.cnki.1007-3639.2023.01.003
  中图分类号： R735.7
- 收稿：2022-09-08，
  
  修回：2022-10-08，
  
  纸质出版：2023-01-30
- 稿件说明：
移动端阅览
胡可舒, 刘文凤, 张锋, 等. 长链非编码RNA LINC00601调控肝细胞癌奥沙利铂耐药的分子机制研究[J]. 中国癌症杂志, 2023,33(1):25-35.

Keshu HU, Wenfeng LIU, Feng ZHANG, et al. Mechanism of LINC00601 in regulating sensitivity of hepatocellular carcinoma cells to oxaliplatin chemotherapy[J]. China Oncology, 2023, 33(1): 25-35.
胡可舒, 刘文凤, 张锋, 等. 长链非编码RNA LINC00601调控肝细胞癌奥沙利铂耐药的分子机制研究[J]. 中国癌症杂志, 2023,33(1):25-35. DOI： 10.19401/j.cnki.1007-3639.2023.01.003.

Keshu HU, Wenfeng LIU, Feng ZHANG, et al. Mechanism of LINC00601 in regulating sensitivity of hepatocellular carcinoma cells to oxaliplatin chemotherapy[J]. China Oncology, 2023, 33(1): 25-35. DOI： 10.19401/j.cnki.1007-3639.2023.01.003.

摘要

背景与目的：

肝细胞癌（hepatocellular carcinoma，HCC）仍是当今中国乃至全世界的高发肿瘤之一，大部分HCC患者确诊时处于中晚期，对化疗耐药或介入治疗抵抗是影响此类患者生存预后的重要因素，但其中涉及的具体分子机制尚未完全明确。我们的前期研究发现长链非编码RNA LINC00601在奥沙利铂耐药的HCC细胞中高表达。本研究旨在探索LINC00601调控HCC细胞对奥沙利铂化疗耐药的作用及其机制，为临床筛选适宜接受奥沙利铂化疗的HCC患者并为中晚期HCC患者的治疗提供新的靶点和理论依据。

方法：

利用体外长期低剂量奥沙利铂刺激人HCC细胞株MHCC-97H和Hep-3B，建立奥沙利铂耐药细胞株97H-OXR和3B-OXR。采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）检测奥沙利铂耐药细胞株中LINC00601的表达情况。进一步采用细胞计数试剂盒-8（cell counting kit-8，CCK-8）实验检测

LINC

00601基因沉默或过表达对HCC细胞奥沙利铂化疗耐药的影响。通过流式细胞术和蛋白质印迹法（Western blot）检测细胞凋亡情况的变化。最后采用Western blot和免疫荧光染色检测生长阻滞和DNA损伤诱导蛋白45α（growth arrest and DNA damage-inducible protein 45α，GADD45A）的核转位，并通过RNA pull-down实验和RNA免疫共沉淀（RNA immunoprecipitation，RIP）实验验证LINC00601和GADD45A的结合。

结果：

与对照组细胞相比，奥沙利铂耐药细胞株中LINC00601的表达水平显著升高。奥沙利铂耐药细胞及过表达LINC00601的MHCC-97H细胞对奥沙利铂体外刺激的敏感性显著下降；相反，下调耐药细胞中LINC00601的表达后，其对于奥沙利铂体外刺激的敏感性明显提高。同时，在奥沙利铂的处理下，高表达LINC00601的HCC细胞株的凋亡水平被明显抑制，且与不同细胞株对奥沙利铂体外处理的敏感性水平相一致。此外，LINC00601的过表达使GADD45A的核转位水平降低，细胞核中GADD45A的表达下降。进一步研究发现LINC00601与GADD45A可以相互结合。

结论：

LINC00601可能通过调控GADD45A的核转位，进而抑制奥沙利铂处理后的细胞凋亡，导致HCC奥沙利铂耐药。

Abstract

Background and purpose:

Hepatocellular carcinoma (HCC) is still one of the most prevalent malignancies in China and throughout the world

while most of these patients are in intermediate or advanced stage when diagnosed. The resistance of advanced HCC to chemotherapy or interventional therapy is one of the crucial factors significantly limiting the survival and prognosis of patients. However

the molecular mechanisms involved have not been fully clarified. We previously found that the long non-coding RNA LINC00601 was highly expressed in oxaliplatin resistant H

CC cells. This study aimed to explore the role and specific mechanism of LINC00601 in regulating the sensitivity of HCC cells to oxaliplatin chemotherapy

to provide a novel target to identify proper candidates for oxaliplatin chemotherapy

and the theoretical basis for the treatment of patients with advanced HCC.

Methods:

Human HCC cell line MHCC-97H and Hep-3B were induced to establish oxaliplatin resistant cell lines 97H-OXR and 3B-OXR by long-term low-dose oxaliplatin treatment

in vitro

. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression of LINC00601 in cell lines. Cell counting kit-8 (CCK-8) assay was performed to detect the effect of LINC00601 gene silencing or overexpression on the sensitivity of HCC cells to oxaliplatin treatment. Flow cytometry and Western blot experiments were conducted to detect the alteration of cell apoptosis. Finally

Western blot and immunofluorescence staining were applied to explore the nuclear translocation of growth arrest and DNA damage-inducible protein 45α (GADD45A)

and RNA pull-down together with RNA immunoprecipitation (RIP) experiments were carried out to validate the combination of LINC00601 and GADD45A.

Results:

Compared with the control cells

LINC00601 expression level was significantly increased in oxaliplatin resistant cells. Both induced-oxaliplatin-resistant cells and LINC00601-overexpressed MHCC-97H cells were observed to be less sensitive to oxaliplatin administration. On the contrary

the down-regulation of LINC00601 expression in oxaliplatin resistant cells significantly increased the sensitivity to oxaliplatin treatment. Meanwhile

upregulated expression of LINC00601 inhibited the apoptosis level of HCC cell lines after oxaliplatin treatment

which was consistent with the sensitivity levels of different cell lines to oxaliplatin as above. In addition

the overexpression of LINC00601 reduced the nuclear translocation of GADD45A and the expression of GADD45A in t

he nucleus. Furthermore

the combination of LINC00601 and GADD45A was observed.

Conclusion:

LINC00601 might regulate the apoptotic progress of cells after oxaliplatin chemotherapy by interfering with the nuclear translocation of GADD45A

which eventually lead to the resistance to oxaliplatin.

关键词

Keywords

references

丁惠国 , 屠红 , 曲春枫 , 等 . 原发性肝癌的分层筛查与监测指南(2020版) [J ] . 临床肝胆病杂志 , 2021 , 37 ( 2 ): 286 - 295 .

DING H G , TU H , QU C F , et al . Guideline for stratified screening and surveillance of primary liver cancer (2020 edition) [J ] . J Clin Hepatol , 2021 , 37 ( 2 ): 286 - 295 .

HU K S , LU S X , LI M , et al . A novel pre-treatment model predicting risk of developing refractoriness to transarterial chemoembolization in unresectable hepatocellular carcinoma [J ] . J Cancer , 2020 , 11 ( 15 ): 4589 - 4596 . DOI: 10.7150/jca.44847 http://doi.org/10.7150/jca.44847

XIE D Y , REN Z G , ZHOU J , et al . 2019 Chinese clinical guidelines for the management of hepatocellular carcinoma: updates and insights [J ] . Hepatobiliary Surg Nutr , 2020 , 9 ( 4 ): 452 - 463 . DOI: 10.21037/hbsn-20-480 http://doi.org/10.21037/hbsn-20-480 http://hbsn.amegroups.com/article/view/47223/html http://hbsn.amegroups.com/article/view/47223/html

SCHMITZ S U , GROTE P , HERRMANN B G . Mechanisms of long noncoding RNA function in development and disease [J ] . Cell Mol Life Sci , 2016 , 73 ( 13 ): 2491 - 2509 . DOI: 10.1007/s00018-016-2174-5 http://doi.org/10.1007/s00018-016-2174-5

YIN X , ZHENG S S , ZHANG L , et al . Identification of long noncoding RNA expression profile in oxaliplatin-resistant hepatocellular carcinoma cells [J ] . Gene , 2017 , 596 : 53 - 88 . DOI: S0378-1119(16)30803-4 http://doi.org/S0378-1119(16)30803-4

SHI X B , LI Y , SUN Y C , et al . Genome-wide analysis of lncRNAs, miRNAs, and mRNAs forming a prognostic scoring system in esophageal squamous cell carcinoma [J ] . PeerJ , 2020 , 8 : e8368 . DOI: 10.7717/peerj.8368 http://doi.org/10.7717/peerj.8368 https://peerj.com/articles/8368 https://peerj.com/articles/8368

HE Y K , YE Y F , TIAN W , et al . A novel lncRNA panel related to ferroptosis, tumor progression, and microenvironment is a robust prognostic indicator for glioma patients [J ] . Front Cell Dev Biol , 2021 , 9 : 788451 . DOI: 10.3389/fcell.2021.788451 http://doi.org/10.3389/fcell.2021.788451 https://www.frontiersin.org/articles/10.3389/fcell.2021.788451/full https://www.frontiersin.org/articles/10.3389/fcell.2021.788451/full

WANG Y C , HU B H , ZHANG W W , et al . Linc00601 upregulation promotes hepatocellular carcinoma development by activating MAPK signaling pathway [J ] . Eur Rev Med Pharmacol Sci , 2020 , 24 ( 11 ): 6039 - 6045 .

RIDDELL I A . Cisplatin and oxaliplatin: our current understanding of their actions [J ] . Met Ions Life Sci , 2018 , 18 : /books/9783110470734/9783110470734-007/9783110470734-007.xml.

曹宇 , 倪娟 , 曹能 , 等 . Gadd45蛋白家族在肿瘤发生、发展中的作用 [J ] . 肿瘤防治研究 , 2019 , 46 ( 3 ): 271 - 274 .

CAO Y , NI J , CAO N , et al . Role of Gadd45 protein family in tumorigenesis and development [J ] . Cancer Res Prev Treat , 2019 , 46 ( 3 ): 271 - 274 .

FENG X M , LIU H , ZHANG Z J , et al . Annexin A2 contributes to cisplatin resistance by activation of JNK-p53 pathway in non-small cell lung cancer cells [J ] . J Exp Clin Cancer Res , 2017 , 36 ( 1 ): 123 . DOI: 10.1186/s13046-017-0594-1 http://doi.org/10.1186/s13046-017-0594-1 http://jeccr.biomedcentral.com/articles/10.1186/s13046-017-0594-1 http://jeccr.biomedcentral.com/articles/10.1186/s13046-017-0594-1

LI J H , XIE X Y , ZHANG L , et al . Oxaliplatin and 5-fluorouracil hepatic infusion with lipiodolized chemoembolization in large hepatocellular carcinoma [J ] . World J Gastroenterol , 2015 , 21 ( 13 ): 3970 - 3977 . DOI: 10.3748/wjg.v21.i13.3970 http://doi.org/10.3748/wjg.v21.i13.3970 http://www.wjgnet.com/1007-9327/full/v21/i13/3970.htm http://www.wjgnet.com/1007-9327/full/v21/i13/3970.htm

GAO S , ZHANG P J , GUO J H , et al . Chemoembolization alone vs combined chemoembolization and hepatic arterial infusion chemotherapy in inoperable hepatocellular carcinoma patients [J ] . World J Gastroenterol , 2015 , 21 ( 36 ): 10443 - 10452 . DOI: 10.3748/wjg.v21.i36.10443 http://doi.org/10.3748/wjg.v21.i36.10443 http://www.wjgnet.com/1007-9327/full/v21/i36/10443.htm http://www.wjgnet.com/1007-9327/full/v21/i36/10443.htm

MARTINEZ-BALIBREA E , MARTÍNEZ-CARDÚS A , GINÉS A , et al . Tumor-related molecular mechanisms of oxaliplatin resistance [J ] . Mol Cancer Ther , 2015 , 14 ( 8 ): 1767 - 1776 . DOI: 10.1158/1535-7163.MCT-14-0636 http://doi.org/10.1158/1535-7163.MCT-14-0636 https://aacrjournals.org/mct/article/14/8/1767/130531/Tumor-Related-Molecular-Mechanisms-of-Oxaliplatin https://aacrjournals.org/mct/article/14/8/1767/130531/Tumor-Related-Molecular-Mechanisms-of-Oxaliplatin

LIM L J , WONG S Y S , HUANG F Y , et al . Roles and regulation of long noncoding RNAs in hepatocellular carcinoma [J ] . Cancer Res , 2019 , 79 ( 20 ): 5131 - 5139 . DOI: 10.1158/0008-5472.CAN-19-0255 http://doi.org/10.1158/0008-5472.CAN-19-0255

HUANG H , CHEN J , DING C M , et al . LncRNA NR2F1-AS1 regulates hepatocellular carcinoma oxaliplatin resistance by targeting ABCC1 via miR-363 [J ] . J Cell Mol Med , 2018 , 22 ( 6 ): 3238 - 3245 . DOI: 10.1111/jcmm.13605 http://doi.org/10.1111/jcmm.13605

HU H Q , YANG L , LI L J , et al . Long non-coding RNA KCNQ1OT1 modulates oxaliplatin resistance in hepatocellular carcinoma through miR-7-5p/ABCC1 axis [J ] . Biochem Biophys Res Commun , 2018 , 503 ( 4 ): 2400 - 2406 . DOI: 10.1016/j.bbrc.2018.06.168 http://doi.org/10.1016/j.bbrc.2018.06.168 https://linkinghub.elsevier.com/retrieve/pii/S0006291X18314888 https://linkinghub.elsevier.com/retrieve/pii/S0006291X18314888

DONG S B , FU Y , YANG K B , et al . Linc01559 served as a potential oncogene and promoted resistance of hepatocellular carcinoma to oxaliplatin by directly sponging miR-6783-3p [J ] . Anticancer Agents Med Chem , 2021 , 21 ( 2 ): 278 - 286 . DOI: 10.2174/1871520620666200721122317 http://doi.org/10.2174/1871520620666200721122317 https://www.eurekaselect.com/183998/article https://www.eurekaselect.com/183998/article

HUANG G L , LI L , LIANG C Y , et al . Upregulated UCA1 contributes to oxaliplatin resistance of hepatocellular carcinoma through inhibition of miR-138-5p and activation of AKT/mTOR signaling pathway [J ] . Pharmacol Res Perspect , 2021 , 9 ( 1 ): e00720 .

XIONG H , NI Z , HE J , et al . LncRNA HULC triggers autophagy via stabilizing Sirt1 and attenuates the chemosensitivity of HCC cells [J ] . Oncogene , 2017 , 36 ( 25 ): 3528 - 3540 . DOI: 10.1038/onc.2016.521 http://doi.org/10.1038/onc.2016.521

ZHU N , SHAO Y , XU L , et al . Gadd45-alpha and Gadd45-gamma utilize p38 and JNK signaling pathways to induce cell cycle G 2 /M arrest in Hep-G2 hepatoma cells [J ] . Mol Biol Rep , 2009 , 36 ( 8 ): 2075 - 2085 . DOI: 10.1007/s11033-008-9419-9 http://doi.org/10.1007/s11033-008-9419-9

TONG T , JI J F , JIN S Q , et al . Gadd45a expression induces Bim dissociation from the cytoskeleton and translocation to mitochondria [J ] . Mol Cell Biol , 2005 , 25 ( 11 ): 4488 - 4500 .

DU K G , LU F , XIE C Z , et al . Toxoplasma gondii infection induces cell apoptosis via multiple pathways revealed by transcriptome analysis [J ] . J Zhejiang Univ Sci B , 2022 , 23 ( 4 ): 315 - 327 . DOI: 10.1631/jzus.B2100877 http://doi.org/10.1631/jzus.B2100877 https://doi.org/10.1631/jzus.B2100877 https://doi.org/10.1631/jzus.B2100877

JANG H J , YANG J H , HONG E , et al . Chelidonine induces apoptosis via GADD45a-p53 regulation in human pancreatic cancer cells [J ] . Integr Cancer Ther , 2021 , 20 : 15347354211006191.

WANG X , ZOU F , ZHONG J T , et al . Secretory clusterin mediates oxaliplatin resistance via the Gadd45a/PI3K/Akt signaling pathway in hepatocellular carcinoma [J ] . J Cancer , 2018 , 9 ( 8 ): 1403 - 1413 . DOI: 10.7150/jca.23849 http://doi.org/10.7150/jca.23849

浏览量

2656

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

长链非编码RNA作为肝细胞癌诊断新型血清标志物的meta分析

PARP1通过调控POU2F2的表达促进肝细胞癌的进展研究

奥沙利铂超敏反应全程管理中国专家共识（2024年版）

M1型肿瘤相关巨噬细胞在肝细胞癌组织中浸润的意义

2022年度肝癌研究及诊疗新进展