Identification of ANXA1 as a Lymphatic Metastasis and Poor Prognostic Factor in Pancreatic Ductal Adenocarcinoma

  • Liu, Qing-Hua (Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute) ;
  • Shi, Mei-Lin (Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute) ;
  • Bai, Jin (Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute) ;
  • Zheng, Jun-Nian (Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute)
  • Published : 2015.04.14


Objective: The aim of this study was to investigate the clinical significance of annexin a1 (ANXA1) and provide molecular evidence to support that decreased ANXA1 expression could enhance cancer migration and invasion in pancreatic ductal adenocarcinoma (PDAC). Materials and Methods: Immunohistochemistry of a tissue microarray with 162 surgically resected PDAC specimens was performed to examine the expression of ANXA1. We also investigated the relationship between ANXA1 expression and clinicopathological factors and prognosis of PDAC patients. We further studied the role of ANXA1 in PDAC cell proliferation, migration and invasion by cell proliferation assay, migration assay and matrigel invasion assay with reduced ANXA1 expression by RNAi. Western blotting was used to detect matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1) expression. We also detected MMP-9 enzyme activity by gelatin zymography. Results: Decreased expression of ANXA1 was significantly associated with poor differentiation, lymph node metastasis and advanced TNM stage of PDAC patients (p<0.05). Moreover, decreased expression of ANXA1 was correlated with poor survival (p<0.05). Furthermore, we found that ANXA1 knockdown inhibited cell proliferation, induced G1 phase cell cycle arrest, increased PDAC cell migration and invasion capacity compared with controls. In addition, Western blotting showed that ANXA1 knockdown increased the MMP-9 protein level and decreased TIMP-1 expression. Gelatin zymography showed that MMP-9 enzyme activity was also elevated. Conclusions: Negative ANXA1 expression is a most unfavorable prognostic factor for PDAC patients. ANXA1 knockdown inhibits cell proliferation by inducing G1 phase cell cycle arrest and increases migration and invasion of PDAC cells through up-regulating MMP-9 expression and activity, implying that ANXA1 may serve as a promising prognostic biomarker and therapeutic target for PDAC.


Annexin a1;pancreatic ductual adenocarcinoma;tissue microarray;lymphatic metastasis;prognostic


Supported by : National Natural Science Foundation of China


  1. Basu B, Correa de Sampaio P, Mohammed H, et al (2012). Inhibition of MT1-MMP activity using functional antibody fragments selected against its hemopexin domain. Int J Biochem Cell Biol, 44, 393-403.
  2. Blackwell GJ, Carnuccio R, Di Rosa M, et al (1980). Macrocortin: a polypeptide causing the anti-phospholipase effect of glucocorticoids. Nature, 287, 147-9.
  3. Cao Y, Li Y, Edelweiss M, et al (2008). Loss of annexin A1 expression in breast cancer progression. Appl Immunohistochem Mol Morphol, 16, 530-4.
  4. Chakraborty A, Dutta J, Das S, et al (2013). Comparison of ex vivo cultivated human limbal epithelial stem cell viability and proliferation on different substrates. Int Ophthalmol, 33, 665-70.
  5. Chao YL, Shepard CR, Wells A (2010). Breast carcinoma cells re-express E-cadherin during mesenchymal to epithelial reverting transition. Mol Cancer, 9, 179.
  6. Chen CY, Shen JQ, Wang F, Wan R, Wang XP (2012). Prognostic significance of annexin A1 expression in pancreatic ductal adenocarcinoma. Asian Pac J Cancer Prev, 13, 4707-12.
  7. Flower RJ, Blackwell GJ (1979). Anti-inflammatory steroids induce biosynthesis of a phospholipase A2 inhibitor which prevents prostaglandin generation. Nature, 278, 456-9.
  8. Hongsrichan N, Rucksaken R, Chamgramol Y, et al (2013). Annexin A1: A new immunohistological marker of cholangiocarcinoma. World J Gastroenterol, 19, 2456-65.
  9. Hynes RO (2003). Metastatic potential: generic predisposition of the primary tumor or rare, metastatic variants-or both? Cell, 113, 821-3.
  10. Inokuchi J, Lau A, Tyson DR, et al (2009). Loss of annexin A1 disrupts normal prostate glandular structure by inducing autocrine IL-6 signaling. Carcinogenesis, 30, 1082-8.
  11. Kang JS, Calvo BF, Maygarden SJ, et al (2002). Dysregulation of annexin I protein expression in high-grade prostatic intraepithelial neoplasia and prostate cancer. Clin Cancer Res, 8, 117-23.
  12. Keleg S, Buchler P, Ludwig R, et al (2003). Invasion and metastasis in pancreatic cancer. Mol Cancer, 2, 14.
  13. Li Y, Wen T, Zhu M, et al (2013). Glycoproteomic analysis of tissues from patients with colon cancer using lectin microarrays and nanoLC-MS/MS. Mol Biosyst, 9, 1877-87.
  14. Lim LH, Pervaiz S (2007). Annexin 1: the new face of an old molecule. FASEB J, 21, 968-75.
  15. Nan Y, Yang S, Tian Y, et al (2009). Analysis of the expression protein profiles of lung squamous carcinoma cell using shot-gun proteomics strategy. Med Oncol, 26, 215-21.
  16. Schittenhelm J, Trautmann K, Tabatabai G, et al (2009). Comparative analysis of annexin-1 in neuroepithelial tumors shows altered expression with the grade of malignancy but is not associated with survival. Mod Pathol, 22, 1600-11.
  17. Siegel R, Naishadham D, Jemal A (2012). Cancer statistics, 2012. CA Cancer J Clin, 62, 10-29.
  18. Swa HL, Shaik AA, Lim LH, et al (2014). Mass spectrometry-based quantitative proteomics and integrative network analysis accentuates modulating roles of Annexin-1 in mammary tumorigenesis. Proteomics.
  19. Wells A, Griffith L, Wells JZ, et al (2013). The dormancy dilemma: quiescence versus balanced proliferation. Cancer Res, 73, 3811-6.
  20. Yates C, Shepard CR, Papworth G, et al (2007). Novel three-dimensional organotypic liver bioreactor to directly visualize early events in metastatic progression. Adv Cancer Res, 97, 225-46.
  21. Yu G, Wang J, Chen Y, et al (2008). Tissue microarray analysis reveals strong clinical evidence for a close association between loss of annexin A1 expression and nodal metastasis in gastric cancer. Clin Exp Metastasis, 25, 695-702.
  22. Zeng GQ, Cheng AL, Tang J, et al (2013). Annexin A1: A new biomarker for predicting nasopharyngeal carcinoma response to radiotherapy. Med Hypotheses, 81, 68-70.

Cited by

  1. The restorative role of annexin A1 at the blood–brain barrier vol.13, pp.1, 2016,
  2. Evaluation of miRNA-196a2 and apoptosis-related target genes: ANXA1, DFFA and PDCD4 expression in gastrointestinal cancer patients: A pilot study vol.12, pp.11, 2017,
  3. iTRAQ-based quantitative proteomic analysis of differentially expressed proteins in chemoresistant nasopharyngeal carcinoma vol.19, pp.9, 2018,