Tumor Biology

, Volume 35, Issue 12, pp 11719–11726 | Cite as

CRTC2 and PROM1 expression in non-small cell lung cancer: analysis by Western blot and immunohistochemistry

  • Yalun Li
  • Yanqi He
  • Zhixin Qiu
  • Bin Zhou
  • Shaoqin Shi
  • Kui Zhang
  • Yangkun Luo
  • Qian Huang
  • Weimin LiEmail author
Research Article


Accumulating evidence supports that genetic factors are another risk factors for lung cancer. Previously, we used whole exome sequencing with sanger sequencing to search for genetic-related mutations in one of four individuals from a pedigree with lung cancer history. Then, we used PCR-RFLP and direct-sequence in the sample size of 318 individuals with lung cancer (cases) and 272 controls. Recently, we detected two new genes including CRTC2 (CREB regulated transcription coactivator 2) and PROM1(human prominin-1,CD133). We investigated the CRTC2 mutation and PROM1 mutation of surgically resected NSCLC tissues (n=200). The presence or absence of CRTC2 and PROM1 mutation was analyzed by direct sequencing. The expression of CRTC2 and PROM1 was studied by western blot and immunohistochemical analysis of the lung cancer tissues which had the mutation of the two genes(cases), the samples without mutations(controls) and the normal lung tissue(controls). CRTC2 and PROM1 mutations in 5 NSCLC tissues and 3 NSCLC tissues out of the samples were identified. The positive results were closely correlated with clinicopathological features, such as male gender, adenocarcinoma, smoker status, and older age (≥55). We found that the CRTC2 and PROM1 expression were significantly higher in tissues of NSCLS with mutations than that without mutations and the normal lung tissue. The results imply that the high expression of CRTC2 and PROM1 may play an important role in the development and hereditary of NSCLC.


CRTC2 PROM1 NSCLC Over-expression 



This study was supported by National Natural Science Foundation of China (grant no. 8120851 to L.D and no. 81241068 to W.L.), Science and technology support program of Sichuan Provincial Science and Technology Department (grant no. 2011SZ0194 to W.L.), and International cooperation program of Sichuan Provincial Science and Technology Department (grant no. 2011HH0051 to W.L.).

Conflicts of interest



  1. 1.
    Alberg AJ, Brock MV, Samet JM. Epidemiology of lung cancer: looking to the future [J]. J Clin Oncol. 2005;23(14):3175–85.PubMedCrossRefGoogle Scholar
  2. 2.
    Lichtenstein P, Holm NV, Verkasalo PK, et al. Environmental and heritable factors in the causation of cancer—analyses of cohorts of twins from Sweden, Denmark, and Finland [J]. N Engl J Med. 2000;343(2):78–85.PubMedCrossRefGoogle Scholar
  3. 3.
    Hedges DJ, Burges D, Powell E, et al. Exome sequencing of a multigenerational human pedigree [J]. PLoS ONE. 2009;4(12):e8232.PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    He Y, Li Y, Qiu Z, et al. Identification and validation of PROM1 and CRTC2 mutations in lung cancer patients [J]. Mol Cancer. 2014;13(1):19.PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Koo SH, Flechner L, Qi L, et al. The CREB coactivator TORC2 is a key regulator of fasting glucose metabolism [J]. Nature. 2005;437(7062):1109–11.PubMedCrossRefGoogle Scholar
  6. 6.
    Mizrak D, Brittan M, Alison M. CD133: molecule of the moment [J]. J Pathol. 2008;214(1):3–9.PubMedCrossRefGoogle Scholar
  7. 7.
    Singh SK, Clarke ID, Terasaki M, et al. Identification of a cancer stem cell in human brain tumors [J]. Cancer Res. 2003;63(18):5821–8.PubMedGoogle Scholar
  8. 8.
    Collins AT, Berry PA, Hyde C, et al. Prospective identification of tumorigenic prostate cancer stem cells [J]. Cancer Res. 2005;65(23):10946–51.PubMedCrossRefGoogle Scholar
  9. 9.
    O’Brien CA, Pollett A, Gallinger S, et al. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice [J]. Nature. 2007;445(7123):106–10.PubMedCrossRefGoogle Scholar
  10. 10.
    Ricci-Vitiani L, Lombardi DG, Pilozzi E, et al. Identification and expansion of human colon-cancer-initiating cells [J]. Nature. 2007;445(7123):111–5.PubMedCrossRefGoogle Scholar
  11. 11.
    Jemal A, Siegel R, Ward E, et al. Cancer statistics, 2008 [J]. CA Cancer J Clin. 2008;58(2):71–96.PubMedCrossRefGoogle Scholar
  12. 12.
    Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics, 2002 [J]. CA Cancer J Clin. 2005;55(2):74–108.PubMedCrossRefGoogle Scholar
  13. 13.
    Sun S, Schiller JH, Gazdar AF. Lung cancer in never smokers—a different disease [J]. Nat Rev Cancer. 2007;7(10):778–90.PubMedCrossRefGoogle Scholar
  14. 14.
    Young RP, Hopkins RJ, Hay BA, et al. Lung cancer susceptibility model based on age, family history and genetic variants [J]. PLoS ONE. 2009;4(4):e5302.PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Gao Y, Goldstein AM, Consonni D, et al. Family history of cancer and nonmalignant lung diseases as risk factors for lung cancer [J]. Int J Cancer. 2009;125(1):146–52.PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Socinski MA, Bogart JA. Limited-stage small-cell lung cancer: the current status of combined-modality therapy [J]. J Clin Oncol. 2007;25(26):4137–45.PubMedCrossRefGoogle Scholar
  17. 17.
    Bernstein ED, Herbert SM, Hanna NH. Chemotherapy and radiotherapy in the treatment of resectable non-small-cell lung cancer [J]. Ann Surg Oncol. 2006;13(3):291–301.PubMedCrossRefGoogle Scholar
  18. 18.
    Poleri C, Morero JL, Nieva B, et al. Risk of recurrence in patients with surgically resected stage I non-small cell lung carcinoma: histopathologic and immunohistochemical analysis [J]. Chest. 2003;123(6):1858–67.PubMedCrossRefGoogle Scholar
  19. 19.
    Screaton RA, Conkright MD, Katoh Y, et al. The CREB coactivator TORC2 functions as a calcium- and cAMP-sensitive coincidence detector [J]. Cell. 2004;119(1):61–74.PubMedCrossRefGoogle Scholar
  20. 20.
    Iourgenko V, Zhang W, Mickanin C, et al. Identification of a family of cAMP response element-binding protein coactivators by genome-scale functional analysis in mammalian cells [J]. Proc Natl Acad Sci U S A. 2003;100(21):12147–52.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Dentin R, Liu Y, Koo SH, et al. Insulin modulates gluconeogenesis by inhibition of the coactivator TORC2 [J]. Nature. 2007;449(7160):366–9.PubMedCrossRefGoogle Scholar
  22. 22.
    Lerner RG, Depatie C, Rutter GA, et al. A role for the CREB co-activator CRTC2 in the hypothalamic mechanisms linking glucose sensing with gene regulation [J]. EMBO Rep. 2009;10(10):1175–81.PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    Fang WL, Lee MT, Wu LS, et al. CREB coactivator CRTC2/TORC2 and its regulator calcineurin crucially mediate follicle-stimulating hormone and transforming growth factor beta1 upregulation of steroidogenesis [J]. J Cell Physiol. 2012;227(6):2430–40.PubMedCrossRefGoogle Scholar
  24. 24.
    Brown KA, Simpson ER. Obesity and breast cancer: progress to understanding the relationship [J]. Cancer Res. 2010;70(1):4–7.PubMedCrossRefGoogle Scholar
  25. 25.
    Pardal R, Clarke MF, Morrison SJ. Applying the principles of stem-cell biology to cancer [J]. Nat Rev Cancer. 2003;3(12):895–902.PubMedCrossRefGoogle Scholar
  26. 26.
    Weigmann A, Corbeil D, Hellwig A, et al. Prominin, a novel microvilli-specific polytopic membrane protein of the apical surface of epithelial cells, is targeted to plasmalemmal protrusions of non-epithelial cells [J]. Proc Natl Acad Sci U S A. 1997;94(23):12425–30.PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Miraglia S, Godfrey W, Yin AH, et al. A novel five-transmembrane hematopoietic stem cell antigen: isolation, characterization, and molecular cloning [J]. Blood. 1997;90(12):5013–21.PubMedGoogle Scholar
  28. 28.
    Fargeas CA, Corbeil D, Huttner WB. AC133 antigen, CD133, prominin-1, prominin-2, etc.: prominin family gene products in need of a rational nomenclature [J]. Stem Cells. 2003;21(4):506–8.PubMedCrossRefGoogle Scholar
  29. 29.
    Singh SK, Hawkins C, Clarke ID, et al. Identification of human brain tumour initiating cells [J]. Nature. 2004;432(7015):396–401.PubMedCrossRefGoogle Scholar
  30. 30.
    Yin S, Li J, Hu C, et al. CD133 positive hepatocellular carcinoma cells possess high capacity for tumorigenicity [J]. Int J Cancer. 2007;120(7):1444–50.PubMedCrossRefGoogle Scholar
  31. 31.
    Olempska M, Eisenach PA, Ammerpohl O, et al. Detection of tumor stem cell markers in pancreatic carcinoma cell lines [J]. Hepatobiliary Pancreat Dis Int. 2007;6(1):92–7.PubMedGoogle Scholar
  32. 32.
    Bruno S, Bussolati B, Grange C, et al. CD133+ renal progenitor cells contribute to tumor angiogenesis [J]. Am J Pathol. 2006;169(6):2223–35.PubMedCentralPubMedCrossRefGoogle Scholar
  33. 33.
    Rutella S, Bonanno G, Procoli A, et al. Cells with characteristics of cancer stem/progenitor cells express the CD133 antigen in human endometrial tumors [J]. Clin Cancer Res. 2009;15(13):4299–311.PubMedCrossRefGoogle Scholar
  34. 34.
    Ferrandina G, Bonanno G, Pierelli L, et al. Expression of CD133-1 and CD133-2 in ovarian cancer [J]. Int J Gynecol Cancer. 2008;18(3):506–14.PubMedCrossRefGoogle Scholar
  35. 35.
    Monzani E, Facchetti F, Galmozzi E, et al. Melanoma contains CD133 and ABCG2 positive cells with enhanced tumourigenic potential [J]. Eur J Cancer. 2007;43(5):935–46.PubMedCrossRefGoogle Scholar
  36. 36.
    Eramo A, Lotti F, Sette G, et al. Identification and expansion of the tumorigenic lung cancer stem cell population [J]. Cell Death Differ. 2008;15(3):504–14.PubMedCrossRefGoogle Scholar
  37. 37.
    Maw MA, Corbeil D, Koch J, et al. A frameshift mutation in prominin (mouse)-like 1 causes human retinal degeneration [J]. Hum Mol Genet. 2000;9(1):27–34.PubMedCrossRefGoogle Scholar
  38. 38.
    Zhang Q, Zulfiqar F, Xiao X, et al. Severe retinitis pigmentosa mapped to 4p15 and associated with a novel mutation in the PROM1 gene [J]. Hum Genet. 2007;122(3–4):293–9.PubMedCrossRefGoogle Scholar
  39. 39.
    Rappa G, Fodstad O, Lorico A. The stem cell-associated antigen CD133 (Prominin-1) is a molecular therapeutic target for metastatic melanoma [J]. Stem Cells. 2008;26(12):3008–17.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Yalun Li
    • 1
  • Yanqi He
    • 1
  • Zhixin Qiu
    • 1
  • Bin Zhou
    • 2
  • Shaoqin Shi
    • 3
  • Kui Zhang
    • 4
  • Yangkun Luo
    • 5
  • Qian Huang
    • 6
  • Weimin Li
    • 1
    Email author
  1. 1.Department of Respiratory MedicineWest China Hospital of Sichuan UniversityChengduChina
  2. 2.Laboratory of Molecular Translational Medicine, West China Institute of Women and Children’s Health, The Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second University HospitalSichuan UniversityChengduChina
  3. 3.Department of Immunology, West China School of Preclinical and Forensic MedicineSichuan UniversityChengduChina
  4. 4.Department of Forensic Biology, West China School of Preclinical and Forensic MedicineSichuan UniversityChengduChina
  5. 5.Department of Radiation OncologyThe Second People’s Hospital of SichuanChengduChina
  6. 6.Department of Clinic LaboratoryChengdu Tumor HospitalChengduChina

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