MutT Homolog 1 (MTH1) maintains multiple KRAS-driven pro-malignant pathways


Oncogenic RAS promotes production of reactive oxygen species (ROS), which mediate pro-malignant signaling but can also trigger DNA damage-induced tumor suppression. Thus RAS-driven tumor cells require redox-protective mechanisms to mitigate the damaging aspects of ROS. Here, we show that MutT Homolog 1 (MTH1), the mammalian 8-oxodGTPase that sanitizes oxidative damage in the nucleotide pool, is important for maintaining several KRAS-driven pro-malignant traits in a nonsmall cell lung carcinoma (NSCLC) model. MTH1 suppression in KRAS-mutant NSCLC cells impairs proliferation and xenograft tumor formation. Furthermore, MTH1 levels modulate KRAS-induced transformation of immortalized lung epithelial cells. MTH1 expression is upregulated by oncogenic KRAS and correlates positively with high KRAS levels in NSCLC human tumors. At a molecular level, in p53-competent KRAS-mutant cells, MTH1 loss provokes DNA damage and induction of oncogene-induced senescence. In p53-nonfunctional KRAS-mutant cells, MTH1 suppression does not produce DNA damage but reduces proliferation and leads to an adaptive decrease in KRAS expression levels. Thus, MTH1 not only enables evasion of oxidative DNA damage and its consequences, but can also function as a molecular rheostat for maintaining oncogene expression at optimal levels. Accordingly, our results indicate MTH1 is a novel and critical component of oncogenic KRAS-associated malignancy and its inhibition is likely to yield significant tumor-suppressive outcomes in KRAS-driven tumors.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5


  1. 1

    Campbell PM, Der CJ Oncogenic Ras and its role in tumor cell invasion and metastasis. Semin Cancer Biol 2004; 14: 105–114.

  2. 2

    Rajalingam K, Schreck R, Rapp UR, Albert S Ras oncogenes and their downstream targets. Biochim Biophys Acta 2007; 1773: 1177–1195.

  3. 3

    Mitsushita J, Lambeth JD, Kamata T The superoxide-generating oxidase Nox1 is functionally required for Ras oncogene transformation. Cancer Res 2004; 64: 3580–3585.

  4. 4

    Arbiser JL, Petros J, Klafter R, Govindajaran B, McLaughlin ER, Brown LF et al. Reactive oxygen generated by Nox1 triggers the angiogenic switch. Proc Natl Acad Sci USA 2002; 99: 715–720.

  5. 5

    Weinberg F, Hamanaka R, Wheaton WW, Weinberg S, Joseph J, Lopez M et al. Mitochondrial metabolism and ROS generation are essential for Kras-mediated tumorigenicity. Proc Natl Acad Sci USA 2010; 107: 8788–8793.

  6. 6

    Irani K, Xia Y, Zweier JL, Sollott SJ, Der CJ, Fearon ER et al. Mitogenic signaling mediated by oxidants in ras-transformed fibroblasts. Science 1997; 275: 1649–1652.

  7. 7

    Moiseeva O, Bourdeau V, Roux A, Deschenes-Simard X, Ferbeyre G Mitochondrial dysfunction contributes to oncogene-induced senescence. Mol Cell Biol 2009; 29: 4495–4507.

  8. 8

    Finkel T Intracellular redox regulation by the family of small GTPases. Antioxid Redox Signal 2006; 8: 1857–1863.

  9. 9

    Rai P, Young JJ, Burton DG, Giribaldi MG, Onder TT, Weinberg RA Enhanced elimination of oxidized guanine nucleotides inhibits oncogenic RAS-induced DNA damage and premature senescence. Oncogene 2011; 30: 1489–1496.

  10. 10

    Mallette FA, Gaumont-Leclerc M-F, Ferbeyre G The DNA damage signaling pathway is a critical mediator of oncogene-induced senescence. Genes Dev 2007; 21: 43–48.

  11. 11

    Di Micco R, Fumagalli M, Cicalese A, Piccinin S, Gasparini P, Luise C et al. Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature 2006; 444: 638–642.

  12. 12

    Liou JS, Chen CY, Chen JS, Faller DV Oncogenic ras mediates apoptosis in response to protein kinase C inhibition through the generation of reactive oxygen species. J Biol Chem 2000; 275: 39001–39011.

  13. 13

    Chen CY, Liou J, Forman LW, Faller DV Correlation of genetic instability and apoptosis in the presence of oncogenic Ki-Ras. Cell Death Differ 1998; 5: 984–995.

  14. 14

    Nakabeppu Y Molecular genetics and structural biology of human MutT homolog, MTH1. Mutat Res 2001; 477: 59–70.

  15. 15

    Rai P, Onder TT, Young JJ, McFaline JL, Pang B, Dedon PC et al. Continuous elimination of oxidized nucleotides is necessary to prevent rapid onset of cellular senescence. Proc Natl Acad Sci USA 2009; 106: 169–174.

  16. 16

    Kennedy CH, Pass HI, Mitchell JB Expression of human MutT homologue (hMTH1) protein in primary non-small-cell lung carcinomas and histologically normal surrounding tissue. Free Radic Biol Med 2003; 34: 1447–1457.

  17. 17

    Cho WC, Chow AS, Au JS MiR-145 inhibits cell proliferation of human lung adenocarcinoma by targeting EGFR and NUDT1. RNA Biol 2011; 8: 125–131.

  18. 18

    Wiederschain D, Wee S, Chen L, Loo A, Yang G, Huang A et al. Single-vector inducible lentiviral RNAi system for oncology target validation. Cell Cycle 2009; 8: 498–504.

  19. 19

    Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 1997; 88: 593–602.

  20. 20

    Collado M, Medema RH, Garcia-Cao I, Dubuisson ML, Barradas M, Glassford J et al. Inhibition of the phosphoinositide 3-kinase pathway induces a senescence-like arrest mediated by p27Kip1. J Biol Chem 2000; 275: 21960–21968.

  21. 21

    Lee AC, Fenster BE, Ito H, Takeda K, Bae NS, Hirai T et al. Ras proteins induce senescence by altering the intracellular levels of reactive oxygen species. J Biol Chem 1999; 274: 7936–7940.

  22. 22

    Takahashi A, Ohtani N, Yamakoshi K, Iida S-i, Tahara H, Nakayama K et al. Mitogenic signalling and the p16INK4a-Rb pathway cooperate to enforce irreversible cellular senescence. Nat Cell Biol 2006; 8: 1291–1297.

  23. 23

    Nogueira V, Park Y, Chen CC, Xu PZ, Chen ML, Tonic I et al. Akt determines replicative senescence and oxidative or oncogenic premature senescence and sensitizes cells to oxidative apoptosis. Cancer Cell 2008; 14: 458–470.

  24. 24

    Janik J, Swoboda M, Janowska B, Ciesla JM, Gackowski D, Kowalewski J et al. 8-Oxoguanine incision activity is impaired in lung tissues of NSCLC patients with the polymorphism of OGG1 and XRCC1 genes. Mutat Res 2011; 709-710: 21–31.

  25. 25

    Wikman H, Risch A, Klimek F, Schmezer P, Spiegelhalder B, Dienemann H et al. hOGG1 polymorphism and loss of heterozygosity (LOH): significance for lung cancer susceptibility in a caucasian population. Int J Cancer 2000; 88: 932–937.

  26. 26

    Speina E, Arczewska KD, Gackowski D, Zielinska M, Siomek A, Kowalewski J et al. Contribution of hMTH1 to the maintenance of 8-oxoguanine levels in lung dna of non-small-cell lung cancer patients. J Natl Cancer Inst 2005; 97: 384–395.

  27. 27

    Dobbs TA, Palmer P, Maniou Z, Lomax ME, O'Neill P Interplay of two major repair pathways in the processing of complex double-strand DNA breaks. DNA Repair (Amst) 2008; 7: 1372–1383.

  28. 28

    Hu CM, Yeh MT, Tsao N, Chen CW, Gao QZ, Chang CY et al. Tumor cells require thymidylate kinase to prevent dUTP incorporation during DNA repair. Cancer Cell 2012; 22: 36–50.

  29. 29

    Liu B, Chen Y St, Clair DK ROS and p53: a versatile partnership. Free Radic Biol Med 2008; 44: 1529–1535.

  30. 30

    Zhou J, Ahn J, Wilson SH, Prives C A role for p53 in base excision repair. EMBO J 2001; 20: 914–923.

  31. 31

    Fan S, el-Deiry WS, Bae I, Freeman J, Jondle D, Bhatia K et al. p53 gene mutations are associated with decreased sensitivity of human lymphoma cells to DNA damaging agents. Cancer Res 1994; 54: 5824–5830.

  32. 32

    Lee JM, Bernstein A p53 mutations increase resistance to ionizing radiation. Proc Natl Acad Sci USA 1993; 90: 5742–5746.

  33. 33

    Hommura F, Dosaka-Akita H, Mishina T, Nishi M, Kojima T, Hiroumi H et al. Prognostic significance of p27KIP1 protein and ki-67 growth fraction in non-small cell lung cancers. Clin Cancer Res 2000; 6: 4073–4081.

  34. 34

    Brummelkamp TR, Bernards R, Agami R Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell 2002; 2: 243–247.

  35. 35

    Sunaga N, Shames DS, Girard L, Peyton M, Larsen JE, Imai H et al. Knockdown of oncogenic KRAS in non-small cell lung cancers suppresses tumor growth and sensitizes tumor cells to targeted therapy. Mol Cancer Ther 2011; 10: 336–346.

  36. 36

    Shirasawa S, Furuse M, Yokoyama N, Sasazuki T Altered growth of human colon cancer cell lines disrupted at activated Ki-ras. Science 1993; 260: 85–88.

  37. 37

    Mukhopadhyay T, Tainsky M, Cavender AC, Roth JA Specific inhibition of K-ras expression and tumorigenicity of lung cancer cells by antisense RNA. Cancer Res 1991; 51: 1744–1748.

  38. 38

    Cao J, Schulte J, Knight A, Leslie NR, Zagozdzon A, Bronson R et al. Prdx1 inhibits tumorigenesis via regulating PTEN/AKT activity. EMBO J 2009; 28: 1505–1517.

  39. 39

    Nogueira V, Hay N Molecular pathways: reactive oxygen species homeostasis in cancer cells and implications for cancer therapy. Clin Cancer Res 2013; 19: 4309–4314.

  40. 40

    Cogoi S, Xodo LE G-quadruplex formation within the promoter of the KRAS proto-oncogene and its effect on transcription. Nucleic Acids Res 2006; 34: 2536–2549.

  41. 41

    Clark DW, Phang T, Edwards MG, Geraci MW, Gillespie MN Promoter G-quadruplex sequences are targets for base oxidation and strand cleavage during hypoxia-induced transcription. Free Radic Biol Med 2012; 53: 51–59.

  42. 42

    Szalai VA, Singer MJ, Thorp HH Site-specific probing of oxidative reactivity and telomerase function using 7,8-dihydro-8-oxoguanine in telomeric DNA. J Am Chem Soc 2002; 124: 1625–1631.

  43. 43

    Ghosh A, Rossi ML, Aulds J, Croteau D, Bohr VA Telomeric D-loops containing 8-oxo-2'-deoxyguanosine are preferred substrates for Werner and Bloom syndrome helicases and are bound by POT1. J Biol Chem 2009; 284: 31074–31084.

  44. 44

    Downward J Targeting RAS signalling pathways in cancer therapy. Nat Rev Cancer 2003; 3: 11–22.

  45. 45

    Huber KV, Salah E, Radic B, Gridling M, Elkins JM, Stukalov A et al. Stereospecific targeting of MTH1 by (S)-crizotinib as an anticancer strategy. Nature 2014; 508: 222–227.

  46. 46

    Gad H, Koolmeister T, Jemth AS, Eshtad S, Jacques SA, Strom CE et al. MTH1 inhibition eradicates cancer by preventing sanitation of the dNTP pool. Nature 2014; 508: 215–221.

  47. 47

    Patel A, Munoz A, Halvorsen K, Rai P Creation and validation of a ligation-independent cloning (LIC) retroviral vector for stable gene transduction in mammalian cells. BMC Biotechnol 2012; 12: 3.

  48. 48

    Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS et al. Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 2003; 9: 493–501.

  49. 49

    Fei DL, Li H, Kozul CD, Black KE, Singh S, Gosse JA et al. Activation of Hedgehog signaling by the environmental toxicant arsenic may contribute to the etiology of arsenic-induced tumors. Cancer Res 2010; 70: 1981–1988.

  50. 50

    Schmittgen TD, Livak KJ Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008; 3: 1101–1108.

  51. 51

    Burton DG, Giribaldi MG, Munoz A, Halvorsen K, Patel A, Jorda M et al. Androgen deprivation-induced senescence promotes outgrowth of androgen-refractory prostate cancer cells. PLoS ONE 2013; 8: e68003.

  52. 52

    Dimri G, Lee X, Basile G, Acosta M, Scott G, Roskelley C et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. PNAS 1995; 92: 9363–9367.

  53. 53

    Reiner T, de las Pozas A, Perez-Stable C Sequential combinations of flavopiridol and docetaxel inhibit prostate tumors, induce apoptosis, and decrease angiogenesis in the Ggamma/T-15 transgenic mouse model of prostate cancer. Prostate 2006; 66: 1487–1497.

Download references


We thank Dr Ramiro Verdun for helpful discussions, and Karen Alvarez Delfin for technical assistance. This work was supported by a James and Esther King Florida Biomedical New Investigator Research grant, a University of Miami Dean’s Bridge Fund award and an NIH/NCI grant (R01CA175086) to PR.

Author information

Correspondence to P Rai.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Supplementary Information (DOC 67 kb)

Supplementary Figure S1 (JPG 797 kb)

Supplementary Figure S2 (JPG 168 kb)

Supplementary Figure S3 (JPG 2534 kb)

Supplementary Figure S4 (JPG 1722 kb)

Supplementary Figure S5 (JPG 1962 kb)

Supplementary Figure S6 (JPG 1196 kb)

Supplementary Figure S7 (JPG 1140 kb)

Supplementary Figure S8 (JPG 1712 kb)

Supplementary Figure S9 (JPG 2068 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Further reading