LMO7 promotes the growth of hepatocellular carcinoma by targeting ferroptosis

doi:10.3877/cma.j.issn.2095-3232.2024.03.019

Abstract

Abstract:

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.

Key words: Carcinoma, hepatocellular, LMO7, Ferroptosis, Cell proliferation, Cell cycle, Reactive oxygen species (ROS)

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.

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Figures/Tables 6

References 23

[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.

引物Primer	引物序列（5'-3'）
hqLMO7-F	GTCTACAGTTCCGTCAAGAAGG
hqLMO7-R	TCTGAAGGATAAGTTGCTCCCT
hqGAPDH-F	AGGTCGGTGTGAACGGATTTG
hqGAPDH-R	TGTAGACCATGTAGTTGAGGTCA

引物Primer	引物序列（5'-3'）
hqLMO7-F	GTCTACAGTTCCGTCAAGAAGG
hqLMO7-R	TCTGAAGGATAAGTTGCTCCCT
hqGAPDH-F	AGGTCGGTGTGAACGGATTTG
hqGAPDH-R	TGTAGACCATGTAGTTGAGGTCA

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