Regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib

CHEN Dahong; WU Yafei; DIAO Wenjing; YANG Huihua; MAO Pengjuan; LI Qin

doi:10.6039/j.issn.1001-0408.2024.08.11

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Regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib

更新时间：2025-09-25

- Regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib
- ZHONGGUO YAOFANG Vol. 35, Issue 8, Pages: 961-966(2024)
- 作者机构：
  
  上海交通大学医学院附属第一人民医院临床药学科，上海　200080
- 作者简介：
- 基金信息：
- DOI：10.6039/j.issn.1001-0408.2024.08.11
  CLC： R965;R735.7
- Received：18 October 2023，
  
  Revised：2024-02-26，
  
  Accepted：11 March 2024，
  
  Published：30 April 2024
- 稿件说明：
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陈大红,吴亚菲,刁文婧,等.自噬在人肝癌细胞Huh7对仑伐替尼耐药中的调节作用[J].中国药房,2024,35(08):961-966.

CHEN Dahong,WU Yafei,DIAO Wenjing,et al.Regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib[J].ZHONGGUO YAOFANG,2024,35(08):961-966.
陈大红,吴亚菲,刁文婧,等.自噬在人肝癌细胞Huh7对仑伐替尼耐药中的调节作用[J].中国药房,2024,35(08):961-966. DOI： 10.6039/j.issn.1001-0408.2024.08.11.

CHEN Dahong,WU Yafei,DIAO Wenjing,et al.Regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib[J].ZHONGGUO YAOFANG,2024,35(08):961-966. DOI： 10.6039/j.issn.1001-0408.2024.08.11.

摘要

目的

研究自噬在人肝癌细胞Huh7对仑伐替尼耐药中的调节作用。

方法

以人肝癌细胞Huh7为对象，采用低浓度逐渐递增联合长期给药的方式构建仑伐替尼耐药细胞模型Huh7-LR，以CCK-8实验和流式细胞术检测亲本细胞Huh7及耐药细胞Huh7-LR对仑伐替尼的敏感性，以Western blot实验和GFP-mCherry-LC3质粒转染实验分别检测细胞中自噬效应蛋白Beclin-1、自噬接头蛋白sequestosome 1（又名p62）、微管相关蛋白1轻链3（LC3）的表达水平和细胞自噬水平。通过饥饿诱导构建自噬激活细胞模型，以Western blot实验和GFP-mCherry-LC3质粒转染实验分别检测细胞中p62蛋白表达水平和细胞自噬水平，以流式细胞术检测自噬激活对仑伐替尼敏感性的影响。

结果

与亲本细胞相比，Huh7-LR细胞的耐药指数为6.2，其p62蛋白的表达水平显著升高，凋亡率、Beclin-1蛋白的表达水平和LC3Ⅱ/LC3Ⅰ比值均显著降低（

＜0.05或

＜0.01），自噬水平有所下降。自噬激活可显著升高Huh7-LR细胞中p62蛋白的表达水平（

＜0.05）和自噬水平，并显著升高其凋亡率（

＜0.05）。

结论

自噬参与了仑伐替尼耐药，激活自噬可一定程度逆转肝癌细胞对仑伐替尼耐药。

Abstract

OBJECTIVE

To investigate the regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib.

METHODS

Using human liver cancer cell Huh7 as subject， the lenvatinib-resist cell model （Huh7-LR） was generated by the low-dose gradient method combined with long-term administration. The sensitivity of parental cell Huh7 and drug-resistant cell Huh7-LR to lenvatinib was detected by using CCK-8 assay and flow cytometry. Western blot assay and GFP-mCherry-LC3 plasmid transfection were performed to detect the expression levels of autophagic protein Beclin-1， autophagic adapter protein sequestosome 1 （p62）， microtubule-associated protein 1 light chain 3 （LC3） and autophagic level. Furthermore， an autophagy activation model was constructed by cell starvation， the protein expression of p62 and autophagy level were detected by using Western blot assay and GFP-mCherry-LC3 plasmid transfection， and the effect of autophagy activation on the sensitivity of Huh7-LR cells to lenvatinib was detected by flow cytometry.

RESULTS

Compared with parental cells， the drug resistance index of Huh7-LR cells was 6.2； protein expression of p62 was increased significantly， while apoptotic rate， protein expression of Beclin-1 and LC3Ⅱ/ LC3Ⅰ ratio were all reduced significantly （

＜0.05 or

＜0.01）； the level of autophagy was decreased to some extent. Autophagy activation could significantly increase the protein expression of p62 in Huh7-LR cells （

＜0.05） and autophagy level， and significantly increase its apoptotic rate （

＜0.05）.

CONCLUSIONS

Autophagy is involved in lenvatinib resistance， and activating autophagy can reverse the resistance of liver cancer cells to lenvatinib to some extent.

关键词

Keywords

references

RUMGAY H ， ARNOLD M ， FERLAY J ， et al . Global burden of primary liver cancer in 2020 and predictions to 2040 [J ] . J Hepatol ， 2022 ， 77 （ 6 ）： 1598 - 1606 .

YANG J D ， HEIMBACH J K . New advances in the diagnosis and management of hepatocellular carcinoma [J ] . BMJ ， 2020 ， 371 ： m3544 .

AL-SALAMA Z T ， SYED Y Y ， SCOTT L J . Lenvatinib：a review in hepatocellular carcinoma [J ] . Drugs ， 2019 ， 79 （ 6 ）： 665 - 674 .

TANG W W ， CHEN Z Y ， ZHANG W L ， et al . The mechanisms of sorafenib resistance in hepatocellular carcinoma：theoretical basis and therapeutic aspects [J ] . Signal Transduct Target Ther ， 2020 ， 5 （ 1 ）： 87 .

CHEN D H ， ZHANG J G ， WU C X ， et al . Non-coding RNA m6A modification in cancer：mechanisms and therapeutic targets [J ] . Front Cell Dev Biol ， 2021 ， 9 ： 778582 .

FU R D ， JIANG S T ， LI J Y ， et al . Activation of the HGF/c-MET axis promotes lenvatinib resistance in hepatocel-lular carcinoma cells with high c-MET expression [J ] . Med Oncol ， 2020 ， 37 （ 4 ）： 24 .

DEVIS-JAUREGUI L ， ERITJA N ， DAVIS M L ， et al . Autophagy in the physiological endometrium and cancer [J ] . Autophagy ， 2021 ， 17 （ 5 ）： 1077 - 1095 .

DEBNATH J ， GAMMOH N ， RYAN K M . Autophagy and autophagy-related pathways in cancer [J ] . Nat Rev Mol Cell Biol ， 2023 ， 24 （ 8 ）： 560 - 575 .

WU Y F ， ZHANG J G ， LI Q . Autophagy，an accomplice or antagonist of drug resistance in HCC [J ] . Cell Death Dis ， 2021 ， 12 （ 3 ）： 266 .

FU X T ， SONG K ， ZHOU J ， et al . Tumor-associated ma-crophages modulate resistance to oxaliplatin via inducing autophagy in hepatocellular carcinoma [J ] . Cancer Cell Int ， 2019 ， 19 ： 71 .

LIN Z Y ， NIU Y ， WAN A ， et al . RNA m 6 A methylation regulates sorafenib resistance in liver cancer through FOXO3-mediated autophagy [J ] . EMBO J ， 2020 ， 39 （ 12 ）： e103181 .

ROBERTS J L ， POKLEPOVIC A ， BOOTH L ， et al . The multi-kinase inhibitor lenvatinib interacts with the HDAC inhibitor entinostat to kill liver cancer cells [J ] . Cell Signal ， 2020 ， 70 ： 109573 .

MA X M ， QIU Y M ， SUN Y L ， et al . NOD2 inhibits tumorigenesis and increases chemosensitivity of hepatocellular carcinoma by targeting AMPK pathway [J ] . Cell Death Dis ， 2020 ， 11 （ 3 ）： 174 .

GAMBARDELLA G ， STAIANO L ， MORETTI M N ， et al . GADD34 is a modulator of autophagy during starvation [J ] . Sci Adv ， 2020 ， 6 （ 39 ）： eabb0205 .

LLOVET J M ， KELLEY R K ， VILLANUEVA A ， et al . Hepatocellular carcinoma [J ] . Nat Rev Dis Primers ， 2021 ， 7 （ 1 ）： 6 .

SHI T T ， IWAMA H ， FUJITA K ， et al . Evaluating the effect of lenvatinib on sorafenib-resistant hepatocellular carcinoma cells [J ] . Int J Mol Sci ， 2021 ， 22 （ 23 ）： 13071 .

JIN H J ， SHI Y P ， LV Y Y ， et al . EGFR activation limits the response of liver cancer to lenvatinib [J ] . Nature ， 2021 ， 595 （ 7869 ）： 730 - 734 .

GALLUZZI L ， GREEN D R . Autophagy-independent functions of the autophagy machinery [J ] . Cell ， 2019 ， 177 （ 7 ）： 1682 - 1699 .

LI X H ， HE S K ， MA B Y . Autophagy and autophagy-related proteins in cancer [J ] . Mol Cancer ， 2020 ， 19 （ 1 ）： 12 .

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