Tumor Biology

, Volume 37, Issue 2, pp 2665–2672 | Cite as

M2 isoform of pyruvate kinase (PKM2) is upregulated in Kazakh’s ESCC and promotes proliferation and migration of ESCC cells

  • Qing Liu
  • Meng Liang
  • Tao Liu
  • Lucine Vuitton
  • Shutao Zheng
  • Xiangpeng Gao
  • Mang Lu
  • Xiuling Li
  • Ilyar Sheyhidin
  • Xiaomei Lu
Original Article

Abstract

The objectives of the present study are to explore role of pyruvate kinase isoenzyme type M2 (PKM2) in progression of Kazakh’s esophageal squamous cell carcinoma (ESCC) in Xinjiang, China, and to clarify mechanism of PKM2 in malignant phenotype. PKM2 expression was examined using immunohistochemistry (IHC) in 101 matched pairs of ESCC and normal adjacent tissues (NATs) and using enzyme-linked immunosorbent assay (ELISA) in 35 serum samples of Kazakh’s ESCC and 8 serum samples of healthy subjects. To investigate mechanism, small interfering RNA (siRNA)-PKM2 was transfected into ESCC cells. Cell migration and invasion were evaluated by wound healing and Transwell assays. Apoptosis and cell cycle were analyzed by flow cytometry (FCM). PKM2 expression was significantly higher in ESCC tissues (77.2 %, 78/101) compared with matched NAT (P = 0.003) and also higher in serum samples of Kazakh’s ESCC patients (78.84 ng/mL) compared with healthy subjects (13.55 ng/mL) (P = 0.001). Patients with overexpression of PKM2 had a poor prognosis (P = 0.032). After knockdown of PKM2, cell proliferation, migration, and invasion were significantly reduced (P = 0.001), apoptosis increased (P = 0.001), and cell cycle was arrested at G1 phase. PKM2 overexpression was significantly correlated with the worse outcome of Kazakh’s ESCC. Furthermore, PKM2 was involved in progression of ESCC by promoting proliferation and suppressing apoptosis, accelerating invasion, and influencing cell cycle. PKM2 could be a potential biomarker for molecular classification of ESCC.

Keywords

ESCC Kazakh PKM2 Malignant phenotype 

Notes

Acknowledgments

This project was supported by the Natural Science Foundation of China (Nos. 81160303, 81260359, 81201891, U1303321) and support from Major Science and Technology Projects of the Xinjiang Uygur Autonomous Region (No. 201430123-1).

Conflicts of interest

None

Supplementary material

13277_2015_4073_MOESM1_ESM.doc (860 kb)
Supplementary figure 1 Specificity of primary antibody against human PKM2 was evaluated using antigen preadsorption test method. A: primary antibody against human PKM2 was diluted at 1:100, followed by incubation with formalin-fixed paraffin-embeded ESCC section. B: primary antibody against human PKM2 was incubated with the synthetic peptide of PKM2 (from human PKM2 ELISA kit) at 50ug/mL, followed by incubation with formalin-fixed paraffin-embeded ESCC section. The magnification was 200×. (DOC 859 kb)
13277_2015_4073_MOESM2_ESM.docx (810 kb)
Supplementary figure 2 Observation of Eca109 cells transfected with siRNA-PKM2 by fluorescence microscopy (Magnification × 100). A: 24 h after transfection with siRNA-PKM2; B. 48 h after transfection with siRNA-PKM2; C. 72 h after transfection with siRNA-PKM2. (DOCX 810 kb)
13277_2015_4073_MOESM3_ESM.docx (52 kb)
Supplementary figure 3 Expression of PKM2 mRNA levels in Eca109 after siRNA-PKM2 transfection. Expression level of PKM2 mRNA in Eca109 was significantly down regulated after transfection with siRNA-PKM2 as compared with negative control and untreated cells. *P < 0.05, **P < 0.01 (DOCX 52 kb)

References

  1. 1.
    Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108.CrossRefPubMedGoogle Scholar
  2. 2.
    Xu Y, Yu X, Chen Q, Mao W. Neoadjuvant versus adjuvant treatment: which one is better for resectable esophageal squamous cell carcinoma? World J Surg Oncol. 2012;10:173–80.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Zhang SW et al. An analysis of incidence and mortality of esophageal cancer in China, 2003–2007. China Cancer. 2012;21:241–7.Google Scholar
  4. 4.
    Lambert R, Hainaut P. The multidisciplinary management of gastrointestinal cancer. Epidemiology of oesophagogastric cancer. Best Pract Res Clin Gastroenterol. 2007;21(6):921–45.CrossRefPubMedGoogle Scholar
  5. 5.
    Zheng ST, Vuitton L, Sheyhidin I, Vuitton DA, Zhang YM, Lu XM. North Western China: a place to learn more on oesophageal cancer. I. Behavioural and environmental risk factors. Eur J Gastroenterol Hepatol. 2010;22(8):917–25.CrossRefPubMedGoogle Scholar
  6. 6.
    Umar SB, Fleischer DE. Esophageal cancer: epidemiology, pathogenesis and prevention. Nat Clin Pract Gastroenterol Hepatol. 2008;5(9):517–26.CrossRefGoogle Scholar
  7. 7.
    Qi YJ, Chao WX, Chiu JF. An overview of esophageal squamous cell carcinoma proteomics. J Proteome. 2012;75:3129–37.CrossRefGoogle Scholar
  8. 8.
    Zhang XH, Liu SQ, Guo CM, Zong JW, Sun MZ. The association of Annexin A2 and cancers. Clin Transl Oncol. 2012;14(9):634–40.CrossRefPubMedGoogle Scholar
  9. 9.
    Liu Z, Feng JG, Tuersun A, et al. Proteomic identification of differentially-expressed proteins in esophageal cancer in three ethnic groups in Xinjiang. Mol Biol Rep. 2011;38(5):3261–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Christofk HR, Vander Heiden MG, Harris MH, et al. The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature. 2008;452(7184):230–3.CrossRefPubMedGoogle Scholar
  11. 11.
    Mazurek S. Pyruvate kinase type M2: a key regulator of the metabolic budget system in tumor cells. Int J Biochem Cell Biol. 2011;43(7):969–80.CrossRefPubMedGoogle Scholar
  12. 12.
    Lee JM, Kohn EC. Proteomics as a guiding tool for more effective personalized therapy. Ann Oncol. 2010;Suppl 7:205–10.Google Scholar
  13. 13.
    Matta A, Ralhan R, Desouza LV, et al. Mass spectrometry-based clinical proteomics: head-and-neck cancer biomarkers and drug-targets discovery. Mass Spectrom Rev. 2010;29(6):945–61.CrossRefPubMedGoogle Scholar
  14. 14.
    Dunn BK, Wagner PD, Anderson D, et al. Molecular markers for early detection. Semin Oncol. 2010;37(3):224–42.CrossRefPubMedGoogle Scholar
  15. 15.
    Minami S, Sato Y, Matsumoto T, et al. Proteomic study of sera from patients with bladder cancer: usefulness of S100A8 and S100A9 proteins. Cancer Genomics Proteomics. 2010;7(4):181–9.PubMedGoogle Scholar
  16. 16.
    Chong PK, Lee H, Loh MC, et al. Upregulation of plasma C9 protein in gastric cancer patients. Proteomics. 2010;10(18):3210–21.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Vander Heiden MG, Locasale JW, Swanson KD, et al. Evidence for an alternative glycolytic pathway in rapidly proliferating cells. Science. 2010;329(5998):1492–9.CrossRefPubMedGoogle Scholar
  18. 18.
    Shi HS, Li D, Zhang J, et al. Silencing of pkm2 increases the efficacy of docetaxel in human lung cancer xenografts in mice. Cancer Sci. 2010;101(6):1447–53.CrossRefPubMedGoogle Scholar
  19. 19.
    Jiang K, He B, Lai L, et al. Cyclosporine A inhibits breast cancer cell growth by down regulating the expression of pyruvate kinase subtype M2. Int J Mol Med. 2012;30(2):302–8.PubMedGoogle Scholar
  20. 20.
    Landt S, Jeschke S, Koeninger A, et al. Tumor-specific correlation of tumor M2 pyruvate kinase in pre-invasive, invasive and recurrent cervical cancer. Anticancer Res. 2010;30(2):375–81.PubMedGoogle Scholar
  21. 21.
    Abdullah M, Rani AA, Simadibrata M, et al. The value of fecal tumor M2 pyruvate kinase as a diagnostic tool for colorectal cancer screening. Acta Med Indones. 2012;44(2):94–9.PubMedGoogle Scholar
  22. 22.
    Eigenbrodt E, Basenau D, Holthusen S, et al. Quantification of tumor type M2 pyruvate kinase (Tu M2-PK) in human carcinomas. Anticancer Res. 1997;17(4B):3153–6.PubMedGoogle Scholar
  23. 23.
    Yuan Y, Guo-Qing P, Yan T, et al. A study of PKM2, PFK-1, and ANT1 expressions in cervical biopsy tissues in China. Med Oncol. 2012;29(4):2904–10.CrossRefPubMedGoogle Scholar
  24. 24.
    Lüftner D, Mesterharm J, Akrivakis C, et al. Tumor type M2 pyruvate kinase expression in advanced breast cancer. Anticancer Res. 2000;20(6D):5077–82.PubMedGoogle Scholar
  25. 25.
    Roigas J, Schulze G, Raytarowski S, et al. Tumor M2 pyruvate kinase in plasma of patients with urological tumors. Tumour Biol. 2001;22(5):282–5.CrossRefPubMedGoogle Scholar
  26. 26.
    Lim JY, Yoon SO, Seol SY, et al. Overexpression of the M2 isoform of pyruvate kinase is an adverse prognostic factor for signet ring cell gastric cancer. World J Gastroenterol. 2012;18(30):4037–43.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Karachaliou N, Papadaki C, Lagoudaki E, et al. Predictive value of BRCA1, ERCC1, ATP7B, PKM2, TOPOI, TOPΟ-IIA, TOPOIIB and C-MYC genes in patients with small cell lung cancer (SCLC) who received first line therapy with cisplatin and etoposide. PLoS One. 2013;8(9):e74611.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Goldberg MS, Sharp PA. Pyruvate kinase M2-specific siRNA induces apoptosis and tumor regression. J Exp Med. 2012;209(2):217–24.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Guo W, Zhang Y, Chen T, et al. Efficacy of RNAi targeting of pyruvate kinase M2 combined with cisplatin in a lung cancer model. J Cancer Res Clin Oncol. 2011;137(1):65–72.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Qing Liu
    • 1
    • 2
  • Meng Liang
    • 1
    • 2
  • Tao Liu
    • 1
    • 2
  • Lucine Vuitton
    • 3
  • Shutao Zheng
    • 1
    • 2
  • Xiangpeng Gao
    • 1
    • 2
  • Mang Lu
    • 1
    • 2
  • Xiuling Li
    • 1
    • 2
  • Ilyar Sheyhidin
    • 2
  • Xiaomei Lu
    • 1
    • 2
  1. 1.Clinical Medical Research Institute, First Affiliated Hospital of Xinjiang Medical UniversityXinjiang Uygur Autonomous RegionUrumqiPeople’s Republic of China
  2. 2.State Key Lab Incubation Base of Xinjiang Major Diseases Research, First Affiliated Hospital of Xinjiang Medical UniversityXinjiang Uygur Autonomous RegionUrumqiPeople’s Republic of China
  3. 3.Department of Gastroenterology, University Hospital Jean MinjozUniversity of Franche-ComteBesançonFrance

Personalised recommendations