切换至 "中华医学电子期刊资源库"

中华肝脏外科手术学电子杂志 ›› 2024, Vol. 13 ›› Issue (03) : 370 -376. doi: 10.3877/cma.j.issn.2095-3232.2024.03.019

基础研究

LMO7通过靶向铁死亡促进肝细胞癌生长
杨兴业1, 彭旭云2, 曾倩1, 梁伟铖2, 肖翠翠3, 郑俊1, 姚嘉1,()   
  1. 1. 510630 广州,中山大学附属第三医院肝脏外科暨肝脏移植中心
    2. 510630 广州,中山大学附属第三医院生物治疗中心
    3. 510630 广州,中山大学附属第三医院麻醉科
  • 收稿日期:2024-02-26 出版日期:2024-06-10
  • 通信作者: 姚嘉
  • 基金资助:
    广东省自然科学基金青年提升项目(2023A1515030052)

LMO7 promotes the growth of hepatocellular carcinoma by targeting ferroptosis

Xingye Yang1, Xuyun Peng2, Qian Zeng1, Weicheng Liang2, Cuicui Xiao3, Jun Zheng1, Jia Yao1,()   

  1. 1. Department of Hepatobiliary Surgery, Liver Transplantation Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
    2. Biological Therapy Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
    3. Department of Anesthesiology, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
  • Received:2024-02-26 Published:2024-06-10
  • Corresponding author: Jia Yao
引用本文:

杨兴业, 彭旭云, 曾倩, 梁伟铖, 肖翠翠, 郑俊, 姚嘉. LMO7通过靶向铁死亡促进肝细胞癌生长[J]. 中华肝脏外科手术学电子杂志, 2024, 13(03): 370-376.

Xingye Yang, Xuyun Peng, Qian Zeng, Weicheng Liang, Cuicui Xiao, Jun Zheng, Jia Yao. LMO7 promotes the growth of hepatocellular carcinoma by targeting ferroptosis[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2024, 13(03): 370-376.

目的

探讨LMO7在肝细胞癌(肝癌)中的表达及其调控肝癌增殖和生长的潜在作用机制。

方法

基于Kaplan-Meier Plotter数据库进行生物信息学分析,验证LMO7与肝癌患者预后的关系。使用shRNA病毒感染后的肝癌细胞HepG2和PLC/PRF/5分成空白对照组(NC)、LMO7敲低组1(shLMO7#1)、LMO7敲低组2(shLMO7#2)3组。采用Western blot和qRT-PCR验证肝癌细胞中LMO7蛋白和mRNA相对表达量,细胞克隆形成实验检测LMO7表达对HepG2细胞增殖能力的影响。采用Western blot检测敲低LMO7后铁死亡相关蛋白ACSL4和P53的表达;流式细胞术检测敲低LMO7后的细胞周期和活性氧(ROS)表达量。两组LMO7表达比较采用t检验,3组比较采用单因素方差分析,组间两两比较采用LSD-t检验。

结果

基于GEO数据库分析显示,肝癌组织LMO7表达明显低于癌旁组织(LSD-t=-3.038,P<0.05)。基于Kaplan-Meier Plotter数据库分析显示,肝癌组织LMO7低表达与患者的不良预后相关(HR=0.50,P<0.05)。细胞克隆形成实验显示,shLMO7#1组和shLMO7#2组平均细胞克隆数分别为(64±10)、(95±26)个,明显多于NC组的(11±5)个(LSD-t= 3.91,6.27;P<0.05)。Western blot检测显示,下调LMO7后肝癌细胞P53蛋白表达显著下调。流式细胞术检测显示,下调LMO7后HepG2细胞主要阻滞在G2/M期;shLMO7#1组和shLMO7#2组LMO7的ROS相对表达量分别为32.6±1.6、47.9±1.0,明显少于NC组的53.3±1.1(LSD-t=-20.12,-5.27;P<0.05)。

结论

LMO7在肝癌患者中低表达,LMO7低表达与患者预后不良有关。低表达LMO7通过靶向抑制铁死亡相关基因P53的表达,进一步调控ROS和细胞周期,从而促进肝癌生长。

Objective

To investigate the expression of LMO7 in hepatocellular carcinoma (HCC) and the potential mechanism of regulating the proliferation and growth of HCC.

Methods

Bioinformatics analysis was carried out based on Kaplan-Meier Plotter database to validate the correlation between LMO7 and prognosis of HCC patients. HepG2 and PLC/PRF/5 HCC cells infected with shRNA virus were divided into 3 groups: normal control group (NC), LMO7 knockdown group 1 (shLMO7#1) and LMO7 knockdown group 2 (SHLMO7#2). The relative expression levels of LMO7 protein and mRNA in HCC cells were determined by Western blot and qRT-PCR. The effect of LMO7 expression on the proliferation of HepG2 cells was assessed by cell colony formation assay. The expression levels of ferroptosis-related proteins ACSL4 and P53 after LMO7 knockdown were detected by Western blot. The cell cycle and expression level of reactive oxygen species (ROS) were assessed by flow cytometry. The expression levels of LMO7 between two groups were compared by t test. The comparison among three groups was performed by one-way ANOVA. Paired comparison between two groups was conducted by LSD-t test.

Results

GEO database analysis showed that the expression of LMO7 in HCC was significantly lower than that in paracancerous tissues (LSD-t=-3.038, P<0.05). Kaplan-Meier Plotter database analysis showed that low expression of LMO7 in HCC tissues was significantly correlated with poor prognosis of HCC patients (HR=0.50, P<0.05). Cell colony formation assay found that the average number of cell colony in the shLMO7#1 and shLMO7#2 groups was 64±10 and 95±26, significantly higher than 11±5 in the NC group (LSD-t=3.91, 6.27; P<0.05). Western blot showed that the expression level of P53 protein in HCC cells was significantly down-regulated after knockdown of LMO7. Flow cytometry showed that HepG2 cells were mainly arrested in G2/Mphase after down-regulation of LMO7. The relative ROS expression levels of LMO7 in the shLMO7#1 and shLMO7#2 groups were 32.6±1.6 and 47.9±1.0, significantly lower than 53.3±1.1 in the NC group (LSD-t=-20.12, -5.27; P<0.05).

Conclusions

LMO7 is lowly expressed in HCC patients, which is associated with poor prognosis of HCC patients. Low expression of LMO7 further regulates ROS and cell cycle through targeted inhibition of ferroptosis-related gene P53, thereby promoting the growth of HCC.

表1 引物序列
图1 LMO7表达水平及其与肝癌患者预后的相关性注:a为GEO数据库分析LMO7在肝癌和癌旁组织中的相对表达量,*为P<0.05;b为Kaplan-Meier Plotter数据库分析LMO7表达与肝癌患者预后的相关性;LMO7为LIM结构域蛋白7,LIHC为肝细胞癌
图2 低表达LMO7肝癌细胞的构建注:a为两种肝癌细胞转染后LMO7 mRNA表达量,b为蛋白表达;NC为空白对照组,shLMO7#为LMO7敲低组;LMO7为LIM结构域蛋白7;*为P<0.05
图3 LMO7对肝癌细胞增殖迁移能力影响的细胞克隆形成实验注:NC为空白对照组,shLMO7#为LMO7敲低组;LMO7为LIM结构域蛋白7;*为P<0.05
图4 肝癌细胞LMO7与铁死亡相关蛋白的关系注:a为凝胶电泳检测显示下调LMO7后P53蛋白表达显著下调;b为GEPIA2数据库分析LMO7基因与铁死亡相关蛋白P53的相关性;NC为空白对照组,shLMO7#为LMO7敲低组;LMO7为LIM结构域蛋白7
图5 流式细胞术检测LMO7低表达对HepG2细胞周期及细胞内ROS的影响注:a为细胞周期,b为ROS;NC为空白对照组,shLMO7#为LMO7敲低组;LMO7为LIM结构域蛋白7,ROS为活性氧;*为P<0.05
[1]
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.
[2]
Anwanwan D, Singh SK, Singh S, et al. Challenges in liver cancer and possible treatment approaches[J]. Biochim Biophys Acta Rev Cancer, 2020, 1873(1):188314.
[3]
Gajos-Michniewicz A, Czyz M. WNT/β-catenin signaling in hepatocellular carcinoma: the aberrant activation, pathogenic roles, and therapeutic opportunities[J]. Genes Dis, 2024, 11(2):727-746.
[4]
Akula SM, Abrams SL, Steelman LS, et al. RAS/RAF/MEK/ERK, PI3K/PTEN/AKT/mTORC1 and TP53 pathways and regulatory miRs as therapeutic targets in hepatocellular carcinoma[J]. Expert Opin Ther Targets, 2019, 23(11):915-929.
[5]
Zhang HF, Gao X, Wang X, et al. The mechanisms of renin-angiotensin system in hepatocellular carcinoma: from the perspective of liver fibrosis, HCC cell proliferation, metastasis and angiogenesis, and corresponding protection measures[J]. Biomedecine Pharmacother, 2021, 141:111868.
[6]
Hu Q, Guo C, Li Y, et al. LMO7 mediates cell-specific activation of the Rho-myocardin-related transcription factor-serum response factor pathway and plays an important role in breast cancer cell migration[J]. Mol Cell Biol, 2011, 31(16):3223-3240.
[7]
Karlsson T, Kvarnbrink S, Holmlund C, et al. LMO7 and LIMCH1 interact with LRIG proteins in lung cancer, with prognostic implications for early-stage disease[J]. Lung Cancer, 2018, 125:174-184.
[8]
He H, Li W, Yan P, et al. Identification of a recurrent LMO7-BRAF fusion in papillary thyroid carcinoma[J]. Thyroid, 2018, 28(6):748-754.
[9]
Hernandez-Gea V, Toffanin S, Friedman SL, et al. Role of the microenvironment in the pathogenesis and treatment of hepatocellular carcinoma[J]. Gastroenterology, 2013, 144(3):512-527.
[10]
Ooshio T, Irie K, Morimoto K, et al. Involvement of LMO7 in the association of two cell-cell adhesion molecules, nectin and E-cadherin, through afadin and alpha-actinin in epithelial cells[J]. J Biol Chem, 2004, 279(30):31365-31373.
[11]
Holaska JM, Rais-Bahrami S, Wilson KL. Lmo7 is an emerin-binding protein that regulates the transcription of emerin and many other muscle-relevant genes[J]. Hum Mol Genet, 2006, 15(23):3459-3472.
[12]
Xie Y, Ostriker AC, Jin Y, et al. LMO7 is a negative feedback regulator of transforming growth factor β signaling and fibrosis[J]. Circulation, 2019, 139(5):679-693.
[13]
Liu X, Yuan H, Zhou J, et al. LMO7 as an unrecognized factor promoting pancreatic cancer progression and metastasis[J]. Front Cell Dev Biol, 2021, 9:647387.
[14]
Takahashi M, Lio CJ, Campeau A, et al. The tumor suppressor kinase DAPK3 drives tumor-intrinsic immunity through the STING-IFN-β pathway[J]. Nat Immunol, 2021, 22(4):485-496.
[15]
Tang D, Chen X, Kang R, et al. Ferroptosis: molecular mechanisms and health implications[J]. Cell Res, 2021, 31(2):107-125.
[16]
Jiang X, Stockwell BR, Conrad M. Ferroptosis: mechanisms, biology and role in disease[J]. Nat Rev Mol Cell Biol, 2021, 22(4):266-282.
[17]
Yao F, Deng Y, Zhao Y, et al. A targetable LIFR-NF-κB-LCN2 axis controls liver tumorigenesis and vulnerability to ferroptosis[J]. Nat Commun, 2021, 12(1):7333.
[18]
Chen J, Li X, Ge C, et al. The multifaceted role of ferroptosis in liver disease[J]. Cell Death Differ, 2022, 29(3):467-480.
[19]
Yang WS, Zeng XF, Liu ZN, et al. Diet and liver cancer risk: a narrative review of epidemiological evidence[J]. Br J Nutr, 2020, 124(3):330-340.
[20]
Huang DQ, Singal AG, Kono Y, et al. Changing global epidemiology of liver cancer from 2010 to 2019: NASH is the fastest growing cause of liver cancer[J]. Cell Metab, 2022, 34(7):969-977, e2.
[21]
Nevola R, Ruocco R, Criscuolo L, et al. Predictors of early and late hepatocellular carcinoma recurrence[J]. World J Gastroenterol, 2023, 29(8):1243-1260.
[22]
莫建涛,杨沛泽,曹瑞奇, 等. 基于生物信息学分析构建肝内胆管细胞癌患者铁死亡相关lncRNA预后模型[J/OL]. 中华肝脏外科手术学电子杂志, 2023, 12(2):185-189.
[23]
Seitz HK, Stickel F. Risk factors and mechanisms of hepatocarcinogenesis with special emphasis on alcohol and oxidative stress[J]. Biol Chem, 2006, 387(4):349-360.
[1] 梁旭, 宋国红. 2023年激素受体阳性/HER-2阴性乳腺癌治疗的研究进展[J]. 中华乳腺病杂志(电子版), 2024, 18(02): 71-77.
[2] 陆朝阳, 金也, 孙备. 腹腔镜解剖性肝切除的发展[J]. 中华普外科手术学杂志(电子版), 2024, 18(04): 363-366.
[3] 曾谣, 谢琴, 陈显育, 王平根, 毛玲秋, 何丹玲, 杜飞, 郑希彦, 何函樨. CDC42EP2基因与肝癌预后、免疫细胞浸润关系及其对细胞迁移侵袭的影响[J]. 中华肝脏外科手术学电子杂志, 2024, 13(03): 363-369.
[4] 钟造茂, 罗文超, 蔡满航, 陈显育, 钟跃思. 肝癌肝切除术后肝衰竭的危险因素分析及列线图模型构建[J]. 中华肝脏外科手术学电子杂志, 2024, 13(03): 289-295.
[5] 王礼光, 严庆, 廖珊, 符荣党, 陈焕伟. 微血管侵犯及手术切缘对肝细胞癌患者术后生存预后的影响[J]. 中华肝脏外科手术学电子杂志, 2024, 13(02): 151-157.
[6] 杨建彬, 陈建华, 张文华, 刘建东. 中心静脉压差值对腹腔镜肝细胞癌肝切除术中出血的影响[J]. 中华肝脏外科手术学电子杂志, 2024, 13(02): 158-162.
[7] 夏辉, 戴斌, 冉君, 王威, 龚昭, 周程. DEP结构域蛋白1B在肝细胞癌中的表达及功能[J]. 中华肝脏外科手术学电子杂志, 2024, 13(02): 205-213.
[8] 陈显育, 曾谣, 莫钊鸿, 翟航, 张广权, 钟造茂, 陈署贤. 生物信息学分析CETP基因在肝癌中表达及其对预后和免疫的影响[J]. 中华肝脏外科手术学电子杂志, 2024, 13(02): 214-219.
[9] 吕垒, 冯啸, 何凯明, 高成立, 杨洲, 贾昌昌, 傅斌生. 组蛋白伴侣VPS72驱动H2AFZ的表达并协同促进肝癌进展[J]. 中华肝脏外科手术学电子杂志, 2024, 13(02): 220-226.
[10] 宋燕京, 乔江春, 宋京海. 中晚期肝癌TACE联合免疫靶向转化治疗后右半肝切除术一例[J]. 中华肝脏外科手术学电子杂志, 2024, 13(02): 227-230.
[11] 朱迎, 赵征, 许达, 陆录, 殷保兵. 免疫检查点抑制剂治疗肝细胞癌的进展与展望[J]. 中华肝脏外科手术学电子杂志, 2024, 13(01): 5-10.
[12] 张占国. 靶向免疫治疗时代的肝癌肝切除术再思考[J]. 中华肝脏外科手术学电子杂志, 2024, 13(01): 11-15.
[13] 孙振, 谭天华, 郑洋洋, 李喆, 宋京海. 基于术前纤维蛋白原与白蛋白比值构建肝癌微血管侵犯的预测模型[J]. 中华肝脏外科手术学电子杂志, 2024, 13(01): 27-32.
[14] 沈佳佳, 何经雄, 王芳, 江艺, 潘凡, 张小进. ICG荧光引导腹腔镜射频消融在合并严重大结节肝硬化小肝癌患者治疗中的应用[J]. 中华肝脏外科手术学电子杂志, 2024, 13(01): 68-71.
[15] 邵文哲, 孙倩男, 王道荣. Galectin-9在结直肠癌中的表达及其临床意义[J]. 中华结直肠疾病电子杂志, 2024, 13(02): 112-120.
阅读次数
全文


摘要