SpringerLink
Log in

Wild-type IDH1 Knockout Leads to G0/G1 Arrest, Impairs Cancer Cell Proliferation, Altering Glycolysis, and the TCA Cycle in Colon Cancer

  • Original Article
  • Published:
Biochemical Genetics Aims and scope Submit manuscript

Abstract

The isocitrate dehydrogenase (IDH), which participates in the TCA cycle, is an important key enzyme in regulating cell metabolism. The effect of the metabolic IDH enzyme on cancer pathogenesis has recently been shown in different types of cancer. However, the role of wild-type (wt) IDH1 in the development of colon cancer is still unknown. Our study investigated the role of the IDH1 enzyme in key hallmarks of colon cancer using various methods such as wound healing, cell cycle, colony formation ability, invasion, and apoptosis analysis. Furthermore, cell metabolism was investigated by pyruvate analysis, dinitrosalicylic acid, and HPLC methods. In addition, CRISPR/Cas9 tool was utilized to knockout the IDH1 gene in colon adenocarcinoma cells (SW620). Further studies were performed in two isogenic IDH1 KO clones. Our findings in both clones suggest that IDH1 KO results in G0/G1 arrest, and reduces proliferation by approximately twofold compared to IDH1 WT cells. In addition, the invasion, migration, and colony formation abilities of IDH1 KO clones were significantly decreased accompanied by significant morphological changes. In the context of metabolism, intracellular glucose, pyruvate, αKG, and malate levels were decreased, while the intracellular citrate level was increased in IDH1 KO clones as compared to IDH1 WT cells. Our results reveal that wt IDH1 knockout leads to a decrease in the aggressive features of colon cancer cells. In conclusion, we reported that wt IDH1 has an effective role in colon cancer progression and could be a potential therapeutic target.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Data Availability

All the generated or analyzed datas during the current study are included in this manuscript. The datas in the present study are available from the corresponding author on reasonable request.

Abbreviations

IDH:

Isocitrate dehydrogenase

wt:

Wild-type

FH:

Fumarate hydratase

SDH:

Succinate dehydrogenase

αKG:

α-Ketoglutarate

MeOH:

Methanol

DNS:

Dinitrosalicylic acid

DNPH:

2,4-Dinitrophenylhydrazine

TBS:

Tris-buffered saline

RT-qPCR:

Reverse transcriptase quantitative polymerase chain reaction

References

  • Alavi M, Rai M (2021) Antisense RNA, the modified CRISPR-Cas9, and metal/metal oxide nanoparticles to inactivate pathogenic bacteria. CMBR 1:52–59

    Google Scholar 

  • Atalay EB, Kayali HA (2022) The elevated D-2-hydroxyglutarate level found as a characteristic metabolic change of colon cancer in both in vitro and in vivo models. Biochem Biophys Res Commun 627:191–199. https://doi.org/10.1016/j.bbrc.2022.08.019

    Article  CAS  PubMed  Google Scholar 

  • Ayar Kayali H, Tarhan L (2006) Vancomycin antibiotic production and TCA-glyoxalate pathways depending on the glucose concentration in Amycolatopsis orientalis. Enzyme Microb Technol 38:727–734

    Article  CAS  Google Scholar 

  • Barnes LD, Kuehn GD, Atkinson DE (1971) Yeast diphosphopyridine nucleotide specific isocitrate dehydrogenase purification and some properties. Biochem 10:3939–3944

    Article  CAS  Google Scholar 

  • Biedermann J, Preussler M, Conde M, Peitzsch M, Richter S, Wiedemuth R (2019) Mutant IDH1 differently affects redox state and metabolism in glial cells of normal and tumor origin. Cancers 11:2028

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bin H, Mei H, Hui W, Bing Z (2022) The correlation between miR -34a-3p, miR -31, PLEK2 and the occurrence, development and prognosis of colorectal cancer. Cell Mol Biol (noisy-Le-Grand) 68:192–200

    Article  PubMed  Google Scholar 

  • Brabetz O, Alla V, Angenendt L, Schliemann C, Berdel WE, Arteaga MF, Mikesch JH (2017) RNA-guided CRISPR-Cas9 system-mediated engineering of acute myeloid leukemia mutations. Mol Ther Nucleic Acids 6:243–248

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cardaci S, Ciriolo MR (2012) TCA cycle defects and cancer: when metabolism tunes redox state. Int J Cell Biol. https://doi.org/10.1155/2012/161837

    Article  PubMed  PubMed Central  Google Scholar 

  • Crooks DR, Maio N, Lang M, Ricketts CJ, Vocke CD, Gurram S, Turan S, Kim YY, Cawthon GM, Sohelian F, Val N, Pfeiffer RM, Jailwala P, Tandon M, Tran B, Fan TMW, Lane AN, Ried T, Wangsa D, Malayeri AA, Merino MJ, Yang Y, Meier JL, Ball MW, Rouault TA, Srinivasan R, Linehan WM (2021) Mitochondrial DNA alterations underlie an irreversible shift to aerobic glycolysis in fumarate hydratase–deficient renal cancer. Sci Signal 14:eabc4436

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dalziel K (1980) Isocitrate dehydrogenase and related oxidative decarboxylases. FEBS Lett 117:45–55

    Article  Google Scholar 

  • Dang L, White DW, Gross S, Bennett BD, Bittinger MA, Driggers EM, Fantin VR, Jang HG, ** S, Keenan MC, Marks KM, Prins RM, Ward PS, Yen KE, Liau LM, Rabinowitz JD, Cantley LC, Thompson CB, Vander Heiden MG, Su SM (2009) Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature 462:739–744

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Franken NA, Rodermond HM, Stap J, Haveman J, Van Bree C (2006) Clonogenic assay of cells in vitro. Nat Protoc 1:2315–2319

    Article  CAS  PubMed  Google Scholar 

  • Hanahan D (2022) Hallmarks of cancer: new dimensions. Cancer Discov 12:31–46

    Article  CAS  PubMed  Google Scholar 

  • Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674

    Article  CAS  PubMed  Google Scholar 

  • Hao HX, Khalimonchuk O, Schraders M, Dephoure N, Bayley JP, Kunst H, Devilee P, Cremers CWRJ, Schiffman JD, Bentz BG, Gygi SP, Winge DR, Kremer H, Rutter J (2009) SDH5, a gene required for flavination of succinate dehydrogenase, is mutated in paraganglioma. Sci 325:1139–1142

    Article  CAS  Google Scholar 

  • Hao J, Wei H, Qi Y, Liu H (2022) miR-129-5p plays an anticancer role in colon cancer by targeting RSF1. Cell Mol Biol 67:196–201

    Article  PubMed  Google Scholar 

  • **ek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier A (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Sci 337:816–821

    Article  CAS  Google Scholar 

  • Khuwijitjaru P, Koomyart I, Kobayashi T, Adachi S (2017) Hydrolysis of konjac flour under subcritical water conditions. Chiang Mai J Sci 44:988–992

    CAS  Google Scholar 

  • Krebs H, Johnson WA (1937) The role of citric acid in intermediate metabolism in animal tissues. Enzymologia 4:148–156

    CAS  Google Scholar 

  • Li L, **aoyu H, Trent JC (2019) Depletion of mutant IDH1 impairs chondrosarcoma growth by downregulating integrins. J Clin Oncol 37:11031–11031

    Article  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Moure CJ, Diplas BH, Chen LH, Yang R, Pirozzi CJ, Wang Z, Spasojevic I, Waitkus MS, He Y, Yan H (2019) CRISPR editing of mutant IDH1 R132H induces a CpG methylation-low state in patient-derived glioma models of G-CIMP. Mol Cancer Res 17:2042–2050

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Neitzel C, Demuth P, Wittmann S, Fahrer J (2020) Targeting altered energy metabolism in colorectal cancer: oncogenic reprogramming, the central role of the TCA cycle and therapeutic opportunities. Cancers 12:1731

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pollard PJ, Ratcliffe PJ (2009) Puzzling Patterns of Predisposition. Sci 324:192–194

    Article  CAS  Google Scholar 

  • Qiao S, Lu W, Glorieux C, Li J, Zeng P, Meng N (2021) Wild-type IDH2 protects nuclear DNA from oxidative damage and is a potential therapeutic target in colorectal cancer. Oncogene 40:5880–5892

    Article  CAS  PubMed  Google Scholar 

  • Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F (2013) Genome engineering using the CRISPR-Cas9 system. Nat Protoc 8:2281–2308

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shi J, Fan L, Li B, Pan H (2022) Molecular Mechanism of Integrin αvβ6 in Liver Metastasis of Colon Cancer Based on SDF-1/CXCR4. Cell Mol Biol 67:88–95

    Article  PubMed  Google Scholar 

  • Sjöblom T, Jones S, Wood LD, Parsons DW, Lin J, Barber TD (2006) The consensus coding sequences of human breast and colorectal cancers. Sci 314:268–274

    Article  Google Scholar 

  • Špačková J, Gotvaldová K, Dvořák A, Urbančoková A, Pospíšilová K, Větvička D, Leguina-Ruzzi A, Tesařová P, Vítek L, Ježek P, Smolková K (2021) Biochemical background in mitochondria affects 2HG production by IDH2 and ADHFE1 in breast carcinoma. Cancers 13:1709

    Article  PubMed  PubMed Central  Google Scholar 

  • Su L, Zhang X, Zheng L, Wang M, Zhu Z, Li P (2020) Mutation of isocitrate dehydrogenase 1 in cholangiocarcinoma impairs tumor progression by inhibiting isocitrate metabolism. Front Endocrinol 11:189

    Article  Google Scholar 

  • Subasi E, Atalay EB, Erdogan D, Sen B, Pakyapan B, Ayar Kayalı H (2020) Synthesis and characterization of thiosemicarbazone-functionalized organoruthenium (II)-arene complexes: Investigation of antitumor characteristics in colorectal cancer cell lines. Mater Sci Eng C 106:110152

    Article  CAS  Google Scholar 

  • Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 71:209–249

    Article  PubMed  Google Scholar 

  • Wang B, Ye Y, Yang X, Liu B, Wang Z, Chen S (2020) SIRT 2-dependent IDH 1 deacetylation inhibits colorectal cancer and liver metastases. EMBO Rep 21:e48183

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wen L, Zhang Y, Yang B, Han F, Ebadi AG, Toughani M (2020) Knockdown of Angiopoietin-like protein 4 suppresses the development of colorectal cancer. Cell Mol Biol 66:117–124

    Article  PubMed  Google Scholar 

  • Wu SS, Li QC, Yin CQ, Xue W, Song CQ (2020) Advances in CRISPR/Cas-based gene therapy in human genetic diseases. Theranostics 10:4374

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • ** Y, Xu P (2021) Global colorectal cancer burden in 2020 and projections to 2040. Transl Oncol 14:101174

    Article  PubMed  PubMed Central  Google Scholar 

  • Yang H, **e S, Liang B, Tang Q, Liu H, Wang D, Huang G (2021) Exosomal IDH1 increases the resistance of colorectal cancer cells to 5-Fluorouracil. J Cancer 12:4862

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yoo KS, Lee EJ, Patil BS (2011) Underestimation of pyruvic acid concentrations by fructose and cysteine in 2, 4-dinitrophenylhydrazine-mediated onion pungency test. J Food Sci 76:C1136–C1142

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We gratefully thank doctoral student Ece Çakıroğlu for her contribution to the CRISPR/Cas9 method.

Funding

The authors declare that there were not any funds, grants, or other support during the preparation of this manuscript.

Author information

Authors and Affiliations

  1. Izmir Biomedicine and Genome Center, Izmir, 35340, Turkey

    Esra Bulut Atalay, Serif Senturk & Hulya Ayar Kayali

  2. Izmir International Biomedicine and Genome Institute (IBG), Dokuz Eylül University, Mithatpasa St. No: 58/5, Balcova, 35340, Izmir, Turkey

    Esra Bulut Atalay, Serif Senturk & Hulya Ayar Kayali

  3. Department of Chemistry, Division of Biochemistry, Faculty of Science, Dokuz Eylul University, 35160, İzmir, Turkey

    Hulya Ayar Kayali

Authors
  1. Esra Bulut Atalay
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Serif Senturk
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hulya Ayar Kayali
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

EBA performed the experiments and analyzed the data and writing- the original draft; SS analyzed the data in the CRISPR/Cas9 method and Writing—review & editing; HAK coordinated all aspects of this work, Writing—review & editing, Supervision HAK contributed to the study conception and design.

Corresponding author

Correspondence to Hulya Ayar Kayali.

Ethics declarations

Conflict of ınterest

There have no relevant financial or non-financial interests by all the authors.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Atalay, E.B., Senturk, S. & Kayali, H.A. Wild-type IDH1 Knockout Leads to G0/G1 Arrest, Impairs Cancer Cell Proliferation, Altering Glycolysis, and the TCA Cycle in Colon Cancer. Biochem Genet 61, 1470–1486 (2023). https://doi.org/10.1007/s10528-022-10325-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10528-022-10325-1

Keywords

Search

Navigation