Advertisement

High calpain-1 expression predicts a poor clinical outcome and contributes to tumor progression in pancreatic cancer patients

  • L. M. Yu
  • Y. S. Zhu
  • C. Z. Xu
  • L. L. Zhou
  • Z. X. Xue
  • Z. Z. Cai
Research Article

Abstract

Background

Pancreatic cancer (PC) is a highly aggressive and metastatic disease, with an elevated mortality rate. It is, therefore, crucial to assess factors affecting the prognosis of PC patients. Meanwhile, calpain-1 is associated with malignant tumor progression and metastasis. Thus, it is meaningful to evaluate the relationship between calpain-1 and PC.

Materials and methods

Calpain-1 protein expression was assessed by immunohistochemistry in 96 pancreatic cancer samples and paired adjacent non-cancerous specimens. In addition, calpain-1 protein levels were assessed in six PC cell lines by western blot (WB). Next, PC cells were transfected with calpain-1 siRNA, and silencing was confirmed by WB. Finally, cell proliferation, colony formation, migration and invasion assays, and cell apoptosis analysis were performed to examine the effects of calpain-1 knockdown on proliferation, growth, apoptosis, migration, and invasion in PC cells.

Results

The results showed that calpain-1 was overexpressed in PC tissues and cells. Meanwhile, calpain-1 overexpression was associated with tumor site (P = 0.029), metastasis (P = 0.000), and TNM stage (P = 0.000), but showed no associations with histological grade (P = 0.396), age (P = 0.809), sex (P = 1.000), and lesion size (P = 0.679). The Kaplan–Meier method demonstrated that the low calpain-1 expression group had increased overall survival (OS) compared with patients expressing high calpain-1 levels (28.7 ± 4.1 vs. 17.0 ± 2.3 months) (P = 0.005). Besides, calpain-1 in PC cells was successfully silenced by liposome-mediated RNA interference, resulting in reduced cell growth, invasion, and metastasis in PC cells, with no effect on apoptosis.

Conclusion

The above findings suggest that calpain-1 should be considered a potential biomarker for PC prognosis and therapy.

Keywords

Pancreatic cancer Calpain-1 Prognosis Proliferation Colony formation Migration Invasion 

Notes

Acknowledgements

This study received research funding from the Zhejiang Provincial Natural Science Foundation of China (Y2100546).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All the experimental procedures have been approved by the ethics committee at The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University.

Informed consent

Resected PC specimens were obtained together with the paired adjacent non-tumor tissue samples from 96 patients at the Second Affiliated Hospital and the First Affiliated Hospital of Wenzhou Medical University, with signed approval obtained from the involved patients.

References

  1. 1.
    Vincent A, Herman J, Schulick R, Hruban RH, Goggins M. Pancreatic cancer. Lancet. 2011;378(9791):607–20.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Wolfgang CL, Herman JM, Laheru DA, et al. Recent progress in pancreatic cancer. CA Cancer J Clin. 2013;63(5):318–48.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kanji ZS, Gallinger S. Diagnosis and management of pancreatic cancer. CMAJ. 2013;185(14):1219–26.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Ryan DP, Hong TS, Bardeesy N. Pancreatic Adenocarcinoma. New Engl J Med. 2014;371(11):1039–49.CrossRefPubMedGoogle Scholar
  5. 5.
    Goll DE, Thompson VF, Li HQ, Wei W, Cong JY. The calpain system. Physiol Rev. 2003;83(3):731–801.CrossRefPubMedGoogle Scholar
  6. 6.
    Storr SJ, Carragher NO, Frame MC, Parr T, Martin SG. The calpain system and cancer. Nat Rev Cancer. 2011;11(5):364–74.CrossRefPubMedGoogle Scholar
  7. 7.
    Potz BA, Abid MR, Sellke FW. Role of calpain in pathogenesis of human disease processes. J Nat Sci. 2016;2(9):e218.PubMedPubMedCentralGoogle Scholar
  8. 8.
    Carragher NO, Frame MC. Calpain: a role in cell transformation and migration. Int J Biochem Cell B. 2002;34(12):1539–43.CrossRefGoogle Scholar
  9. 9.
    Reichrath J, Welter C, Mitschele T, et al. Different expression patterns of calpain isozymes 1 and 2 (CAPN1 and 2) in squamous cell carcinomas (SCC) and basal cell carcinomas (BCC) of human skin. J Pathol. 2003;199(4):509–16.CrossRefPubMedGoogle Scholar
  10. 10.
    Leloup L, Wells A. Calpains as potential anti-cancer targets. Expert Opin Ther Tar. 2011;15(3):309–23.CrossRefGoogle Scholar
  11. 11.
    Ma D, Fang J, Liu Y, et al. High level of calpain1 promotes cancer cell invasion and migration in oral squamous cell carcinoma. Oncol Lett. 2017;13(6):4017–26.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Al-Bahlani SM, Al-Rashdi RM, Kumar S, Al-Sinawi SS, Al-Bahri MA, Shalaby AA. Calpain-1 expression in triple-negative breast cancer: a potential prognostic factor independent of the proliferative/apoptotic index. Biomed Res Int. 2017;2017:9290425.PubMedPubMedCentralGoogle Scholar
  13. 13.
    Luo W, Ren Z, Gao S, et al. Clinical correlation of calpain-1 and glypican-3 expression with gallbladder carcinoma. Oncol Lett. 2016;11(2):1345–52.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Yoshida M, Miyasaka Y, Ohuchida K, et al. Calpain inhibitor calpeptin suppresses pancreatic cancer by disrupting cancer-stromal interactions in a mouse xenograft model. Cancer Sci. 2016;107(10):1443–52.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Guroff G. A neutral, calcium-activated proteinase from the soluble fraction of rat brain. J Biol Chem. 1964;239:149–55.PubMedGoogle Scholar
  16. 16.
    Huttenlocher A, Palecek SP, Lu Q, et al. Regulation of cell migration by the calcium-dependent protease calpain. J Biol Chem. 1997;272(52):32719–22.CrossRefPubMedGoogle Scholar
  17. 17.
    Braun C, Engel M, Seifert M, et al. Expression of calpain I messenger RNA in human renal cell carcinoma: correlation with lymph node metastasis and histological type. Int J Cancer. 1999;84(1):6–9.CrossRefPubMedGoogle Scholar
  18. 18.
    Starska K, Forma E, Jozwiak P, et al. Gene/protein expression of CAPN1/2-CAST system members is associated with ERK1/2 kinases activity as well as progression and clinical outcome in human laryngeal cancer. Tumor Biol. 2016;37(10):13185–203.CrossRefGoogle Scholar
  19. 19.
    Liu BD, Zhou Y, Lu D, et al. Comparison of the protein expression of calpain-1, calpain-2, calpastatin and calmodulin between gastric cancer and normal gastric mucosa. Oncol Lett. 2017;14(3):3705–10.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Storr SJ, Pu X, Davis J, et al. Expression of the calpain system is associated with poor clinical outcome in gastro-oesophageal adenocarcinomas. J Gastroenterol. 2013;48(11):1213–21.CrossRefPubMedGoogle Scholar
  21. 21.
    Demarchi F, Schneider C. The calpain system as a modulator of stress/damage response. Cell Cycle. 2007;6(2):136–8.CrossRefPubMedGoogle Scholar
  22. 22.
    Burrows F, Zhang H, Kamal A. Hsp90 activation and cell cycle regulation. Cell Cycle. 2004;3(12):1530–6.CrossRefPubMedGoogle Scholar
  23. 23.
    Mlynarczuk-Bialy I, Roeckmann H, Kuckelkorn U, et al. Combined effect of proteasome and calpain inhibition on cisplatin-resistant human melanoma cells. Cancer Res. 2006;66(15):7598–605.CrossRefPubMedGoogle Scholar
  24. 24.
    Kassis J, Radinsky R, Wells A. Motility is rate-limiting for invasion of bladder carcinoma cell lines. Int J Biochem Cell B. 2002;34(7):762–75.CrossRefGoogle Scholar

Copyright information

© Federación de Sociedades Españolas de Oncología (FESEO) 2018

Authors and Affiliations

  1. 1.Department of GastroenterologyThe Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical UniversityWenzhouChina
  2. 2.Department of PathologyThe Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical UniversityWenzhouChina

Personalised recommendations