MT1G 通过调控 KLF4/HIF1α/CA9 轴抑制肝癌的发展

  索拉非尼作为一种多靶点激酶抑制剂，已被广泛用作晚期肝癌的一线抗癌药物，可以将肝癌患者的中位生存期延长 3-5 个月。然而，在临床应用中，索拉非尼治疗的患者经常出现耐药现象，从而导致索拉非尼疗效欠佳。

  通过转录组学测序，初步结果发现 MT1G 可以被索拉非尼所激活。本研究以肝癌细胞系和实验小鼠为模型 ，应用多种生物学技术 ，以期阐明MT1G 在抑制肝癌进展中的生物学作用及分子机制。首先，我们在肝癌细胞系中过表达或者敲降 MT1G，探寻 MT1G 在索拉非尼发挥药效中的抗肿瘤作用。此外，为进一步研究 MT1G 的抗肿瘤作用分子机制，我们进行了转录组学测序。根据转录组学测序结果，应用Western Blot、 qPCR、双荧光素酶报告系统、多聚核糖体分析技术、染色质免疫共沉淀等技术进一步研
究下游分子的调控机制。

  结果显示, 索拉非尼通过激活 MT1G 发挥其抗肿瘤作用。 MT1G 表达量与患者预后显著相关，因此其有望成为新的肝癌标志物。 MT1G 过表达抑制肝癌的细胞增殖和肿瘤形成，并使细胞对索拉非尼治疗敏感性增强； 敲降 MT1G 后索拉菲尼药效减弱，表明 MT1G 介导了索拉非尼对肝癌细胞的杀伤作用。 最后，我们发现索拉非尼诱导 MT1G 可以激活转录因子 KLF4，进一步抑制下游 CA9 的表达。 KLF4 可直接结合 CA9 的启动子区域或者抑制 HIF1α 的翻译， 从而抑制 CA9 的转录。 总之， 我们的研究结果表明，MT1G 可以通过激活 KLF4 来调节 HIF1α/CA9 轴抑制肝癌的发展。

[1] SIEGEL R L, MILLER K D, FUCHS H E, et al. Cancer statistics, 2022 [J]. CA Cancer J Clin, 2022, 72(1): 7-33.
[2] TANG W, CHEN Z, ZHANG W, et al. The mechanisms of sorafenib resistance in hepatocellular carcinoma: theoretical basis and therapeutic aspects [J]. Signal Transduct Target Ther, 2020, 5(1): 87.
[3] WANG Y, WANG G, TAN X, et al. MT1G serves as a tumor suppressor in hepatocellular carcinoma by interacting with p53 [J]. Oncogenesis, 2019, 8(12): 67.
[4] HOUESSINON A, FRANCOIS C, SAUZAY C, et al. Metallothionein-1 as a biomarker of altered redox metabolism in hepatocellular carcinoma cells exposed to sorafenib [J]. Mol Cancer, 2016, 15(1): 38.
[5] SUN X, NIU X, CHEN R, et al. Metallothionein-1G facilitates sorafenib resistance through inhibition of ferroptosis [J]. Hepatology, 2016, 64(2): 488-500.
[6] LLOVET J M, KELLEY R K, VILLANUEVA A, et al. Hepatocellular carcinoma [J]. Nat Rev Dis Primers, 2021, 7(1): 6.
[7] ZHENG R, QU C, ZHANG S, et al. Liver cancer incidence and mortality in China: Temporal trends and projections to 2030 [J]. Chin J Cancer Res, 2018, 30(6): 571-9.
[8] EL-SERAG H B, RUDOLPH K L. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis [J]. Gastroenterology, 2007, 132(7): 2557-76.
[9] EUROPEAN ASSOCIATION FOR THE STUDY OF THE L, EUROPEAN ORGANISATION FOR R, TREATMENT OF C. EASL-EORTC clinical practice guidelines: management of hepatocellular carcinoma [J]. J Hepatol, 2012, 56(4): 908-43.
[10] TRINCHET J C, BOURCIER V, CHAFFAUT C, et al. Complications and competing risks of death in compensated viral cirrhosis (ANRS CO12 CirVir prospective cohort) [J]. Hepatology, 2015, 62(3): 737-50.
[11] FRACANZANI A L, CONTE D, FRAQUELLI M, et al. Increased cancer risk in a cohort of 230 patients with hereditary hemochromatosis in comparison to matched control patients with non-iron-related chronic liver disease [J]. Hepatology, 2001, 33(3): 647-51.
[12] CHEN C J, YANG H I, SU J, et al. Risk of hepatocellular carcinoma across a biological gradient of serum hepatitis B virus DNA level [J]. JAMA, 2006, 295(1): 65-73.
[13] MURAKAMI Y, SAIGO K, TAKASHIMA H, et al. Large scaled analysis of hepatitis B virus (HBV) DNA integration in HBV related hepatocellular carcinomas [J]. Gut, 2005, 54(8): 1162-8.
[14] SHIH W L, KUO M L, CHUANG S E, et al. Hepatitis B virus X protein inhibits transforming growth factor-beta -induced apoptosis through the activation of phosphatidylinositol 3-kinase pathway [J]. J Biol Chem, 2000, 275(33): 25858-64.
[15] CHIANG C J, YANG Y W, YOU S L, et al. Thirty-year outcomes of the national hepatitis B immunization program in Taiwan [J]. JAMA, 2013, 310(9): 974-6.
[16] POLARIS OBSERVATORY H C V C. Global prevalence and genotype distribution of hepatitis C virus infection in 2015: a modelling study [J]. Lancet Gastroenterol Hepatol, 2017, 2(3): 161-76.
[17] GAO X, ZHAN M, WANG L, et al. Timing of DAA Initiation After Curative Treatment and Its Relationship with the Recurrence of HCV-Related HCC [J]. J Hepatocell Carcinoma, 2020, 7(347-60.
[18] MUNAKA M, KOHSHI K, KAWAMOTO T, et al. Genetic polymorphisms of tobacco- and alcohol-related metabolizing enzymes and the risk of hepatocellular carcinoma [J]. J Cancer Res Clin Oncol, 2003, 129(6): 355-60.
[19] MCCLAIN C J, HILL D B, SONG Z, et al. Monocyte activation in alcoholic liver disease [J]. Alcohol, 2002, 27(1): 53-61.
[20] LIN C W, LIN C C, MO L R, et al. Heavy alcohol consumption increases the incidence of hepatocellular carcinoma in hepatitis B virus-related cirrhosis [J]. J Hepatol, 2013, 58(4): 730-5.
[21] HUANG D Q, EL-SERAG H B, LOOMBA R. Global epidemiology of NAFLD-related HCC: trends, predictions, risk factors and prevention [J]. Nat Rev Gastroenterol Hepatol, 2021, 18(4): 223-38.
[22] ANSTEE Q M, REEVES H L, KOTSILITI E, et al. From NASH to HCC: current concepts and future challenges [J]. Nat Rev Gastroenterol Hepatol, 2019, 16(7): 411-28.
[23] RICH N E, YOPP A C, SINGAL A G, et al. Hepatocellular Carcinoma Incidence Is Decreasing Among Younger Adults in the United States [J]. Clin Gastroenterol Hepatol, 2020, 18(1): 242-8 e5.
[24] LEE Y C, COHET C, YANG Y C, et al. Meta-analysis of epidemiologic studies on cigarette smoking and liver cancer [J]. Int J Epidemiol, 2009, 38(6): 1497-511.
[25] THORGEIRSSON S S, GRISHAM J W. Molecular pathogenesis of human hepatocellular carcinoma [J]. Nat Genet, 2002, 31(4): 339-46.
[26] GARRIDO A, DJOUDER N. Cirrhosis: A Questioned Risk Factor for Hepatocellular Carcinoma [J]. Trends Cancer, 2021, 7(1): 29-36.
[27] WULFING C, VAN AHLEN H, ELTZE E, et al. Metallothionein in bladder cancer: correlation of overexpression with poor outcome after chemotherapy [J]. World J Urol, 2007, 25(2): 199-205.
[28] BRUIX J, BOIX L, SALA M, et al. Focus on hepatocellular carcinoma [J]. Cancer Cell, 2004, 5(3): 215-9.
[29] WU Y, ZHANG J, ZHANG X, et al. Cancer Stem Cells: A Potential Breakthrough in HCC-Targeted Therapy [J]. Front Pharmacol, 2020, 11(198.
[30] BATLLE E, CLEVERS H. Cancer stem cells revisited [J]. Nat Med, 2017, 23(10): 1124-34.
[31] WAN P T, GARNETT M J, ROE S M, et al. Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF [J]. Cell, 2004, 116(6): 855-67.
[32] CHEN L, LIU S, TAO Y. Regulating tumor suppressor genes: post-translational modifications [J]. Signal Transduct Target Ther, 2020, 5(1): 90.
[33] MENDOZA P R, GROSSNIKLAUS H E. The Biology of Retinoblastoma [J]. Prog Mol Biol Transl Sci, 2015, 134(503-16.
[34] CHATTERJEE N, WALKER G C. Mechanisms of DNA damage, repair, and mutagenesis [J]. Environ Mol Mutagen, 2017, 58(5): 235-63.
[35] SAVAGE K I, HARKIN D P. BRCA1, a 'complex' protein involved in the maintenance of genomic stability [J]. FEBS J, 2015, 282(4): 630-46.
[36] BOXER L M, DANG C V. Translocations involving c-myc and c-myc function [J]. Oncogene, 2001, 20(40): 5595-610.
[37] MITELMAN F, JOHANSSON B, MERTENS F. The impact of translocations and gene fusions on cancer causation [J]. Nat Rev Cancer, 2007, 7(4): 233-45.
[38] CHIANG D Y, VILLANUEVA A, HOSHIDA Y, et al. Focal gains of VEGFA and molecular classification of hepatocellular carcinoma [J]. Cancer Res, 2008, 68(16): 6779-88.
[39] SAWEY E T, CHANRION M, CAI C, et al. Identification of a therapeutic strategy targeting amplified FGF19 in liver cancer by Oncogenomic screening [J]. Cancer Cell, 2011, 19(3): 347-58.
[40] ZUCMAN-ROSSI J, VILLANUEVA A, NAULT J C, et al. Genetic Landscape and Biomarkers of Hepatocellular Carcinoma [J]. Gastroenterology, 2015, 149(5): 1226-39 e4.
[41] HE S, TANG S. WNT/beta-catenin signaling in the development of liver cancers [J]. Biomed Pharmacother, 2020, 132(110851.
[42] HAN T S, BAN H S, HUR K, et al. The Epigenetic Regulation of HCC Metastasis [J]. Int J Mol Sci, 2018, 19(12):
[43] XIA H, OOI L L, HUI K M. MicroRNA-216a/217-induced epithelial-mesenchymal transition targets PTEN and SMAD7 to promote drug resistance and recurrence of liver cancer [J]. Hepatology, 2013, 58(2): 629-41.
[44] YU M, XUE H, WANG Y, et al. miR-345 inhibits tumor metastasis and EMT by targeting IRF1-mediated mTOR/STAT3/AKT pathway in hepatocellular carcinoma [J]. Int J Oncol, 2017, 50(3): 975-83.
[45] SINGAL R, GINDER G D. DNA methylation [J]. Blood, 1999, 93(12): 4059-70.
[46] GONZALO S, BLASCO M A. Role of Rb family in the epigenetic definition of chromatin [J]. Cell Cycle, 2005, 4(6): 752-5.
[47] SAITO Y, KANAI Y, SAKAMOTO M, et al. Expression of mRNA for DNA methyltransferases and methyl-CpG-binding proteins and DNA methylation status on CpG islands and pericentromeric satellite regions during human hepatocarcinogenesis [J]. Hepatology, 2001, 33(3): 561-8.
[48] DONG Y, WANG A. Aberrant DNA methylation in hepatocellular carcinoma tumor suppression (Review) [J]. Oncol Lett, 2014, 8(3): 963-8.
[49] GAO X, SHENG Y, YANG J, et al. Osteopontin alters DNA methylation through up-regulating DNMT1 and sensitizes CD133+/CD44+ cancer stem cells to 5 azacytidine in hepatocellular carcinoma [J]. J Exp Clin Cancer Res, 2018, 37(1): 179.
[50] HAMATSU T, RIKIMARU T, YAMASHITA Y, et al. The role of MTA1 gene expression in human hepatocellular carcinoma [J]. Oncol Rep, 2003, 10(3): 599-604.
[51] LEE M H, NA H, KIM E J, et al. Poly(ADP-ribosyl)ation of p53 induces gene-specific transcriptional repression of MTA1 [J]. Oncogene, 2012, 31(49): 5099-107.
[52] LEE M H, NA H, NA T Y, et al. Epigenetic control of metastasis-associated protein 1 gene expression by hepatitis B virus X protein during hepatocarcinogenesis [J]. Oncogenesis, 2012, 1(e25.
[53] FULLGRABE J, KAVANAGH E, JOSEPH B. Histone onco-modifications [J]. Oncogene, 2011, 30(31): 3391-403.
[54] LU H, LI G, ZHOU C, et al. Regulation and role of post-translational modifications of enhancer of zeste homologue 2 in cancer development [J]. Am J Cancer Res, 2016, 6(12): 2737-54.
[55] XIAO G, JIN L L, LIU C Q, et al. EZH2 negatively regulates PD-L1 expression in hepatocellular carcinoma [J]. J Immunother Cancer, 2019, 7(1): 300.
[56] LUNDBERG L E, STENBERG P, LARSSON J. HP1a, Su(var)3-9, SETDB1 and POF stimulate or repress gene expression depending on genomic position, gene length and expression pattern in Drosophila melanogaster [J]. Nucleic Acids Res, 2013, 41(8): 4481-94.
[57] SHAO Y, SONG X, JIANG W, et al. MicroRNA-621 Acts as a Tumor Radiosensitizer by Directly Targeting SETDB1 in Hepatocellular Carcinoma [J]. Mol Ther, 2019, 27(2): 355-64.
[58] WU H, YANG T Y, LI Y, et al. Tumor Necrosis Factor Receptor-Associated Factor 6 Promotes Hepatocarcinogenesis by Interacting With Histone Deacetylase 3 to Enhance c-Myc Gene Expression and Protein Stability [J]. Hepatology, 2020, 71(1): 148-63.
[59] QU N, BO X, LI B, et al. Role of N6-Methyladenosine (m(6)A) Methylation Regulators in Hepatocellular Carcinoma [J]. Front Oncol, 2021, 11(755206.
[60] CHEN M, WEI L, LAW C T, et al. RNA N6-methyladenosine methyltransferase-like 3 promotes liver cancer progression through YTHDF2-dependent posttranscriptional silencing of SOCS2 [J]. Hepatology, 2018, 67(6): 2254-70.
[61] QIAO K, LIU Y, XU Z, et al. RNA m6A methylation promotes the formation of vasculogenic mimicry in hepatocellular carcinoma via Hippo pathway [J]. Angiogenesis, 2021, 24(1): 83-96.
[62] ZHONG L, LIAO D, ZHANG M, et al. YTHDF2 suppresses cell proliferation and growth via destabilizing the EGFR mRNA in hepatocellular carcinoma [J]. Cancer Lett, 2019, 442(252-61.
[63] SUHAIL Y, CAIN M P, VANAJA K, et al. Systems Biology of Cancer Metastasis [J]. Cell Syst, 2019, 9(2): 109-27.
[64] FIDLER I J. The pathogenesis of cancer metastasis: the 'seed and soil' hypothesis revisited [J]. Nat Rev Cancer, 2003, 3(6): 453-8.
[65] HARPER K L, SOSA M S, ENTENBERG D, et al. Mechanism of early dissemination and metastasis in Her2(+) mammary cancer [J]. Nature, 2016, 540(7634): 588-92.
[66] LEHMANN S, TE BOEKHORST V, ODENTHAL J, et al. Hypoxia Induces a HIF-1-Dependent Transition from Collective-to-Amoeboid Dissemination in Epithelial Cancer Cells [J]. Curr Biol, 2017, 27(3): 392-400.
[67] COLEGIO O R, CHU N Q, SZABO A L, et al. Functional polarization of tumour-associated macrophages by tumour-derived lactic acid [J]. Nature, 2014, 513(7519): 559-63.
[68] MAJIDPOOR J, MORTEZAEE K. Steps in metastasis: an updated review [J]. Med Oncol, 2021, 38(1): 3.
[69] LO H C, XU Z, KIM I S, et al. Resistance to natural killer cell immunosurveillance confers a selective advantage to polyclonal metastasis [J]. Nat Cancer, 2020, 1(7): 709-22.
[70] DE BOCK K, MAZZONE M, CARMELIET P. Antiangiogenic therapy, hypoxia, and metastasis: risky liaisons, or not? [J]. Nat Rev Clin Oncol, 2011, 8(7): 393-404.
[71] WU X, ZHANG X, YU L, et al. Zinc finger protein 367 promotes metastasis by inhibiting the Hippo pathway in breast cancer [J]. Oncogene, 2020, 39(12): 2568-82.
[72] MASSAGUE J, GANESH K. Metastasis-Initiating Cells and Ecosystems [J]. Cancer Discov, 2021, 11(4): 971-94.
[73] DIEPENBRUCK M, CHRISTOFORI G. Epithelial-mesenchymal transition (EMT) and metastasis: yes, no, maybe? [J]. Curr Opin Cell Biol, 2016, 43(7-13.
[74] TUNG-PING POON R, FAN S T, WONG J. Risk factors, prevention, and management of postoperative recurrence after resection of hepatocellular carcinoma [J]. Ann Surg, 2000, 232(1): 10-24.
[75] ZHU Y J, ZHENG B, WANG H Y, et al. New knowledge of the mechanisms of sorafenib resistance in liver cancer [J]. Acta Pharmacol Sin, 2017, 38(5): 614-22.
[76] HAMPTON T. Cancer drug trials show modest benefit: drugs target liver, gastric, head and neck cancers [J]. JAMA, 2007, 298(3): 273-5.
[77] ABOU-ALFA G K, JOHNSON P, KNOX J J, et al. Doxorubicin plus sorafenib vs doxorubicin alone in patients with advanced hepatocellular carcinoma: a randomized trial [J]. JAMA, 2010, 304(19): 2154-60.
[78] AWADA A, GIL T, WHENHAM N, et al. Safety and pharmacokinetics of sorafenib combined with capecitabine in patients with advanced solid tumors: results of a phase 1 trial [J]. J Clin Pharmacol, 2011, 51(12): 1674-84.
[79] ZHU K, HUANG J, LAI L, et al. Medium or Large Hepatocellular Carcinoma: Sorafenib Combined with Transarterial Chemoembolization and Radiofrequency Ablation [J]. Radiology, 2018, 288(1): 300-7.
[80] BLIVET-VAN EGGELPOEL M J, CHETTOUH H, FARTOUX L, et al. Epidermal growth factor receptor and HER-3 restrict cell response to sorafenib in hepatocellular carcinoma cells [J]. J Hepatol, 2012, 57(1): 108-15.
[81] GEDALY R, ANGULO P, HUNDLEY J, et al. PI-103 and sorafenib inhibit hepatocellular carcinoma cell proliferation by blocking Ras/Raf/MAPK and PI3K/AKT/mTOR pathways [J]. Anticancer Res, 2010, 30(12): 4951-8.
[82] ZHAI B, HU F, JIANG X, et al. Inhibition of Akt reverses the acquired resistance to sorafenib by switching protective autophagy to autophagic cell death in hepatocellular carcinoma [J]. Mol Cancer Ther, 2014, 13(6): 1589-98.
[83] SINGH S S, VATS S, CHIA A Y, et al. Dual role of autophagy in hallmarks of cancer [J]. Oncogene, 2018, 37(9): 1142-58.
[84] SHIMIZU S, TAKEHARA T, HIKITA H, et al. Inhibition of autophagy potentiates the antitumor effect of the multikinase inhibitor sorafenib in hepatocellular carcinoma [J]. Int J Cancer, 2012, 131(3): 548-57.
[85] TREDAN O, GALMARINI C M, PATEL K, et al. Drug resistance and the solid tumor microenvironment [J]. J Natl Cancer Inst, 2007, 99(19): 1441-54.
[86] LIANG Y, ZHENG T, SONG R, et al. Hypoxia-mediated sorafenib resistance can be overcome by EF24 through Von Hippel-Lindau tumor suppressor-dependent HIF-1alpha inhibition in hepatocellular carcinoma [J]. Hepatology, 2013, 57(5): 1847-57.
[87] PARK C Y, TSENG D, WEISSMAN I L. Cancer stem cell-directed therapies: recent data from the laboratory and clinic [J]. Mol Ther, 2009, 17(2): 219-30.
[88] FERNANDO J, MALFETTONE A, CEPEDA E B, et al. A mesenchymal-like phenotype and expression of CD44 predict lack of apoptotic response to sorafenib in liver tumor cells [J]. Int J Cancer, 2015, 136(4): E161-72.
[89] MCKEOWN S R. Defining normoxia, physoxia and hypoxia in tumours-implications for treatment response [J]. Br J Radiol, 2014, 87(1035): 20130676.
[90] RIEDL C C, BRADER P, ZANZONICO P B, et al. Imaging hypoxia in orthotopic rat liver tumors with iodine 124-labeled iodoazomycin galactopyranoside PET [J]. Radiology, 2008, 248(2): 561-70.
[91] WADA H, NAGANO H, YAMAMOTO H, et al. Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma: importance of angiopoietin-2 and hypoxia-induced factor-1 alpha [J]. Liver Int, 2006, 26(4): 414-23.
[92] HELMLINGER G, YUAN F, DELLIAN M, et al. Interstitial pH and pO2 gradients in solid tumors in vivo: high-resolution measurements reveal a lack of correlation [J]. Nat Med, 1997, 3(2): 177-82.
[93] SUN X, JIANG H, JIANG X, et al. Antisense hypoxia-inducible factor-1alpha augments transcatheter arterial embolization in the treatment of hepatocellular carcinomas in rats [J]. Hum Gene Ther, 2009, 20(4): 314-24.
[94] BAO M H, WONG C C. Hypoxia, Metabolic Reprogramming, and Drug Resistance in Liver Cancer [J]. Cells, 2021, 10(7):
[95] SEMENZA G L. Hypoxia-inducible factors in physiology and medicine [J]. Cell, 2012, 148(3): 399-408.
[96] CORIAT R, NICCO C, CHEREAU C, et al. Sorafenib-induced hepatocellular carcinoma cell death depends on reactive oxygen species production in vitro and in vivo [J]. Mol Cancer Ther, 2012, 11(10): 2284-93.
[97] YOU L, WU W, WANG X, et al. The role of hypoxia-inducible factor 1 in tumor immune evasion [J]. Med Res Rev, 2021, 41(3): 1622-43.
[98] PASTOREKOVA S, PARKKILA S, PARKKILA A K, et al. Carbonic anhydrase IX, MN/CA IX: analysis of stomach complementary DNA sequence and expression in human and rat alimentary tracts [J]. Gastroenterology, 1997, 112(2): 398-408.
[99] PASTOREK J, PASTOREKOVA S. Hypoxia-induced carbonic anhydrase IX as a target for cancer therapy: from biology to clinical use [J]. Semin Cancer Biol, 2015, 31(52-64.
[100] COYLE P, PHILCOX J C, CAREY L C, et al. Metallothionein: the multipurpose protein [J]. Cell Mol Life Sci, 2002, 59(4): 627-47.
[101] FERRARIO C, LAVAGNI P, GARIBOLDI M, et al. Metallothionein 1G acts as an oncosupressor in papillary thyroid carcinoma [J]. Lab Invest, 2008, 88(5): 474-81.
[102] KUMARI M V, HIRAMATSU M, EBADI M. Free radical scavenging actions of metallothionein isoforms I and II [J]. Free Radic Res, 1998, 29(2): 93-101.
[103] ZHENG Y, JIANG L, HU Y, et al. Metallothionein 1H (MT1H) functions as a tumor suppressor in hepatocellular carcinoma through regulating Wnt/beta-catenin signaling pathway [J]. BMC Cancer, 2017, 17(1): 161.
[104] DEMIDENKO R, DANIUNAITE K, BAKAVICIUS A, et al. Decreased expression of MT1E is a potential biomarker of prostate cancer progression [J]. Oncotarget, 2017, 8(37): 61709-18.
[105] ARRIAGA J M, LEVY E M, BRAVO A I, et al. Metallothionein expression in colorectal cancer: relevance of different isoforms for tumor progression and patient survival [J]. Hum Pathol, 2012, 43(2): 197-208.
[106] RUTTKAY-NEDECKY B, NEJDL L, GUMULEC J, et al. The role of metallothionein in oxidative stress [J]. Int J Mol Sci, 2013, 14(3): 6044-66.
[107] KLAASSEN C D, LIU J, DIWAN B A. Metallothionein protection of cadmium toxicity [J]. Toxicol Appl Pharmacol, 2009, 238(3): 215-20.
[108] LIM D, JOCELYN K M, YIP G W, et al. Silencing the Metallothionein-2A gene inhibits cell cycle progression from G1- to S-phase involving ATM and cdc25A signaling in breast cancer cells [J]. Cancer Lett, 2009, 276(1): 109-17.
[109] WERYNSKA B, PULA B, MUSZCZYNSKA-BERNHARD B, et al. Correlation between expression of metallothionein and expression of Ki-67 and MCM-2 proliferation markers in non-small cell lung cancer [J]. Anticancer Res, 2011, 31(9): 2833-9.
[110] OSTRAKHOVITCH E A, OLSSON P E, JIANG S, et al. Interaction of metallothionein with tumor suppressor p53 protein [J]. FEBS Lett, 2006, 580(5): 1235-8.
[111] KIM H G, KIM J Y, HAN E H, et al. Metallothionein-2A overexpression increases the expression of matrix metalloproteinase-9 and invasion of breast cancer cells [J]. FEBS Lett, 2011, 585(2): 421-8.
[112] YAN D W, FAN J W, YU Z H, et al. Downregulation of metallothionein 1F, a putative oncosuppressor, by loss of heterozygosity in colon cancer tissue [J]. Biochim Biophys Acta, 2012, 1822(6): 918-26.
[113] KIM H G, HWANG Y P, JEONG H G. Metallothionein-III induces HIF-1alpha-mediated VEGF expression in brain endothelial cells [J]. Biochem Biophys Res Commun, 2008, 369(2): 666-71.
[114] SUBRAMANIAN VIGNESH K, DEEPE G S, JR. Metallothioneins: Emerging Modulators in Immunity and Infection [J]. Int J Mol Sci, 2017, 18(10):
[115] EMRI E, EGERVARI K, VARVOLGYI T, et al. Correlation among metallothionein expression, intratumoural macrophage infiltration and the risk of metastasis in human cutaneous malignant melanoma [J]. J Eur Acad Dermatol Venereol, 2013, 27(3): e320-7.
[116] WEST A K, STALLINGS R, HILDEBRAND C E, et al. Human metallothionein genes: structure of the functional locus at 16q13 [J]. Genomics, 1990, 8(3): 513-8.
[117] HIRAKO N, NAKANO H, TAKAHASHI S. A PU.1 suppressive target gene, metallothionein 1G, inhibits retinoic acid-induced NB4 cell differentiation [J]. PLoS One, 2014, 9(7): e103282.
[118] ARRIAGA J M, BRAVO A I, MORDOH J, et al. Metallothionein 1G promotes the differentiation of HT-29 human colorectal cancer cells [J]. Oncol Rep, 2017, 37(5): 2633-51.
[119] FU J, LV H, GUAN H, et al. Metallothionein 1G functions as a tumor suppressor in thyroid cancer through modulating the PI3K/Akt signaling pathway [J]. BMC Cancer, 2013, 13(462.
[120] JI X F, FAN Y C, GAO S, et al. MT1M and MT1G promoter methylation as biomarkers for hepatocellular carcinoma [J]. World J Gastroenterol, 2014, 20(16): 4723-9.
[121] ARRIAGA J M, GRECO A, MORDOH J, et al. Metallothionein 1G and zinc sensitize human colorectal cancer cells to chemotherapy [J]. Mol Cancer Ther, 2014, 13(5): 1369-81.
[122] FARRUGIA M K, VANDERBILT D B, SALKENI M A, et al. Kruppel-like Pluripotency Factors as Modulators of Cancer Cell Therapeutic Responses [J]. Cancer Res, 2016, 76(7): 1677-82.
[123] TETREAULT M P, YANG Y, KATZ J P. Kruppel-like factors in cancer [J]. Nat Rev Cancer, 2013, 13(10): 701-13.
[124] YANG H, PARK D, RYU J, et al. USP11 degrades KLF4 via its deubiquitinase activity in liver diseases [J]. J Cell Mol Med, 2021, 25(14): 6976-87.
[125] LU X J, SHI Y, CHEN J L, et al. Kruppel-like factors in hepatocellular carcinoma [J]. Tumour Biol, 2015, 36(2): 533-41.
[126] SUN H, TANG H, XIE D, et al. Kruppel-like Factor 4 Blocks Hepatocellular Carcinoma Dedifferentiation and Progression through Activation of Hepatocyte Nuclear Factor-6 [J]. Clin Cancer Res, 2016, 22(2): 502-12.
[127] SUN H, PENG Z, TANG H, et al. Loss of KLF4 and consequential downregulation of Smad7 exacerbate oncogenic TGF-beta signaling in and promote progression of hepatocellular carcinoma [J]. Oncogene, 2017, 36(21): 2957-68.
[128] WEI D, KANAI M, HUANG S, et al. Emerging role of KLF4 in human gastrointestinal cancer [J]. Carcinogenesis, 2006, 27(1): 23-31.
[129] OPAVSKY R, PASTOREKOVA S, ZELNIK V, et al. Human MN/CA9 gene, a novel member of the carbonic anhydrase family: structure and exon to protein domain relationships [J]. Genomics, 1996, 33(3): 480-7.
[130] NAKAGAWA Y, UEMURA H, HIRAO Y, et al. Radiation hybrid mapping of the human MN/CA9 locus to chromosome band 9p12-p13 [J]. Genomics, 1998, 53(1): 118-9.
[131] MAHON B P, PINARD M A, MCKENNA R. Targeting carbonic anhydrase IX activity and expression [J]. Molecules, 2015, 20(2): 2323-48.
[132] SUPURAN C T, WINUM J Y. Carbonic anhydrase IX inhibitors in cancer therapy: an update [J]. Future Med Chem, 2015, 7(11): 1407-14.
[133] THIRY A, DOGNE J M, MASEREEL B, et al. Targeting tumor-associated carbonic anhydrase IX in cancer therapy [J]. Trends Pharmacol Sci, 2006, 27(11): 566-73.
[134] PASTOREK J, PASTOREKOVA S, CALLEBAUT I, et al. Cloning and characterization of MN, a human tumor-associated protein with a domain homologous to carbonic anhydrase and a putative helix-loop-helix DNA binding segment [J]. Oncogene, 1994, 9(10): 2877-88.
[135] LYKO F. The DNA methyltransferase family: a versatile toolkit for epigenetic regulation [J]. Nat Rev Genet, 2018, 19(2): 81-92.
[136] YANG J, LIU L, LI M, et al. Naringenin inhibits proinflammatory cytokine production in macrophages through inducing MT1G to suppress the activation of NFkappaB [J]. Mol Immunol, 2021, 137(155-62.
[137] LI K, ZHANG Z, MEI Y, et al. Metallothionein-1G suppresses pancreatic cancer cell stemness by limiting activin A secretion via NF-kappaB inhibition [J]. Theranostics, 2021, 11(7): 3196-212.
[138] ZHU X, DU J, YU J, et al. LncRNA NKILA regulates endothelium inflammation by controlling a NF-kappaB/KLF4 positive feedback loop [J]. J Mol Cell Cardiol, 2019, 126(60-9.
[139] AKAOGI K, NAKAJIMA Y, ITO I, et al. KLF4 suppresses estrogen-dependent breast cancer growth by inhibiting the transcriptional activity of ERalpha [J]. Oncogene, 2009, 28(32): 2894-902.
[140] YASUDA M, HATANAKA T, SHIRATO H, et al. Cell type-specific reciprocal regulation of HIF1A gene expression is dependent on 5'- and 3'-UTRs [J]. Biochem Biophys Res Commun, 2014, 447(4): 638-43.
[141] YANG N, WANG T, LI Q, et al. HBXIP drives metabolic reprogramming in hepatocellular carcinoma cells via METTL3-mediated m6A modification of HIF-1alpha [J]. J Cell Physiol, 2021, 236(5): 3863-80.