Parasitology Research

, Volume 102, Issue 3, pp 411–417 | Cite as

Proliferative effects of excretory/secretory products from Clonorchis sinensis on the human epithelial cell line HEK293 via regulation of the transcription factor E2F1

  • Young Ju Kim
  • Min-Ho Choi
  • Sung-Tae Hong
  • Young Mee Bae
Original Paper


Clonorchis sinensis is one of the most prevalent parasitic helminths of humans in East Asia. Although several complications in bile duct epithelial cells are caused by C. sinensis infection, the mechanism is not clearly understood. To clarify the effects of C. sinensis excretory–secretory products (ES products) on bile duct epithelial cells, we investigated their effects on the human embryonic kidney epithelial cell line HEK293 in vitro. Our results show that ES products alter the proportion of cells in each stage of the cell cycle and induce HEK293 cell proliferation. Among cell cycle-related proteins, the expression of cyclin E increased markedly after treatment with ES products, indicating that the G1/S transition occurred. In addition, the expression of the transcription factor E2F1 was up-regulated by the addition of ES products. Small interfering RNA (siRNA) was used to demonstrate that the transcription factor E2F1 is a key factor in the control of cell proliferation in HEK293 cells. The present results demonstrate that ES products from C. sinensis stimulate cell proliferation by inducing E2F1 expression. We suggest that the ES products released from C. sinensis during infection may play an important role in the development of cholangiocarcinoma via the overgrowth of the bile duct epithelium.


Cell Cycle Progression HEK293 Cell Cholangiocarcinoma Cell Cycle Protein Clonorchiasis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Grant sponsor: Seoul National University Hospital Research Fund; Grant number: 04-2007-051.


  1. Blake MC, Azizkhan JC (1989) Transcription factor E2F1 is required for efficient expression of the hamster dihydrofolate reductase gene in vitro and in vivo. Mol Cell Biol 9:4994–5002PubMedGoogle Scholar
  2. Carmona C, Dowd AJ, Smith AM, Dalton JP (1993) Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles. Mol Biochem Parasitol 62:9–17PubMedCrossRefGoogle Scholar
  3. Cho PY, Lee MJ, Kim TI, Kang SY, Hong SJ (2006) Expressed sequence tag analysis of adult Clonorchis sinensis, the Chinese live fluke. Parasitol Res 99:602–608PubMedCrossRefGoogle Scholar
  4. Choi D, Lim JH, Lee KT, Lee JK, Choi SH, Heo JS, Jang KT, Lee NY, Kim S, Hong ST (2006) Cholangiocarcinoma and clonorchis sinensis infection: a case-control study in Korea. J Hepatol 44:1066–1073PubMedCrossRefGoogle Scholar
  5. Choi MH, Ryu JS, Lee M, Li S, Chung BS, Chai JY, Sithithaworn P, Tesana S, Hong ST (2003) Specific and common antigens of Clonorchis sinensis and Opisthorchis viverrini (Opisthorchidae, Trematoda). Korean J Parasitol 41:155–163PubMedGoogle Scholar
  6. Chung YB, Kong Y, Joo IJ, Cho SY, Kang SY (1995) Excystment of Paragonimus westermani metacercariae by endogenous cysteine protease. J Parasitol 81:137–142PubMedCrossRefGoogle Scholar
  7. Fry CJ, Pearson A, Malinowski E, Bartley SM, Greenblatt J, Farnham PJ (1999) Activation of the murine dihydrofolate reductase promoter by E2F1. J Biol Chem 274:15883–15891PubMedCrossRefGoogle Scholar
  8. Geng Y, Eaton EN, Picón M, Roberts JM, Lundberg AS, Gifford A, Sardet C, Weinberg RA (1996) Regulation of cyclin E transcription by E2Fs and retinoblastoma proteins. Oncogene 12:1173–1180PubMedGoogle Scholar
  9. Goto Y, Hayashi R, Kang D, Yoshida K (2006) Acute loss of transcription factor E2F1 induces mitochondrial biogenesis in HeLa cells. J Cell Physiol 209:923–934PubMedCrossRefGoogle Scholar
  10. Herrera LA, Benítez-Bribiesca L, Mohar A, Ostrosky-Wegman P (2005) Role of infectious diseases in human carcinogenesis. Environ Mol Mutagen 45:284–303PubMedCrossRefGoogle Scholar
  11. Hong ST (2003) Clonorchis sinensis. In: Miliotis MD, Bier JW (eds) International handbook of foodborne pathogens. Marcel Dekker, New York, pp 581–592Google Scholar
  12. Hong ST, Kho WG, Kim WH, Chai JY, Lee SH (1993) Turnover of biliary epithelial cells in Clonorchis sinensis infected rats. Korean J Parasitol 31:83–89PubMedGoogle Scholar
  13. Jin Y, Lee JC, Choi IY, Kim EA, Shin MH, Kim WK (2006) Excretory–secretory products produced by Paragonimus westermani differentially regulate the nitric oxide production and viability of microglial cells. Int Arch Allergy Immunol 139:16–24PubMedCrossRefGoogle Scholar
  14. Johnson DG, Schneider-Broussard R (1998) Role of E2F in cell cycle control and cancer. Front Biosci 3:d447–d448PubMedGoogle Scholar
  15. King KL, Cidlowski JA (1998) Cell cycle regulation and apoptosis. Annu Rev Physiol 60:601–617PubMedCrossRefGoogle Scholar
  16. Kong Y, Chung YB, Cho SY, Kang SY (1994) Cleavage of immunoglobulin G by excretory-secretory cathepsin S-like protease of Spirometra mansoni plerocercoid. Parasitology 109(Pt 5):611–621PubMedCrossRefGoogle Scholar
  17. Kuo YC, Kuo PL, Hsu YL, Cho CY, Lin CC (2006) Ellipticine induces apoptosis through p53-dependent pathway in human hepatocellular carcinoma HepG2 cells. Life Sci 78:2550–2557PubMedCrossRefGoogle Scholar
  18. Kuzmenok OI, Chiang SC, Lin YC, Lee ST (2005) Retardation of cell cycle progression of macrophages from G1 to S phase by ICAM-L from Leishmania. Int J Parasitol 35:1547–1555PubMedCrossRefGoogle Scholar
  19. Lee JH, Rim HJ, Bak UB (1993) Effect of Clonorchis sinensis infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian golden hamsters. Korean J Parasitol 31:21–30PubMedGoogle Scholar
  20. Lee JH, Rim HJ, Sell S (1997) Heterogeneity of the “oval-cell” response in the hamster liver during cholangiocarcinogenesis following Clonorchis sinensis infection and dimethylnitrosamine treatment. J Hepatol 26:1313–1323PubMedCrossRefGoogle Scholar
  21. Lee JH, Yang HM, Bak UB, Rim HJ (1994) Promoting role of Clonorchis sinensis infection on induction of cholangiocarcinoma during two-step carcinogenesis. Korean J Parasitol 32:13–18PubMedCrossRefGoogle Scholar
  22. Lim MK, Ju YH, Franceschi S, Oh JK, Kong HJ, Hwang SS, Park SK, Cho SI, Shon WM, Kim DI, Yoo KY, Hong ST, Shin HR (2006) Clonochis sinensis infection and increasing risk of cholangiocarcinoma in the Republic of Korea. Am J Trop Med Hyg 75:93–96PubMedGoogle Scholar
  23. Martens JS, Reiner NE, Herrera-Velit P, Steinbrecher UP (1998) Phosphatidylinositol 3-kinase is involved in the induction of macrophage growth by oxidized low density lipoprotein. J Biol Chem 273:4915–4920PubMedCrossRefGoogle Scholar
  24. Matsumura I, Tanaka H, Kanakura Y (2003) E2F1 and c-Myc in cell growth and death. Cell Cycle 2:333–338PubMedGoogle Scholar
  25. Mundle SD, Saberwal G (2003) Evolving intricacies and implications of E2F1 regulation. FASEB J 17:569–574PubMedCrossRefGoogle Scholar
  26. Nordeen SK (1988) Luciferase reporter gene vectors for analysis promoters and enhancers. Biotechniques 6:454–458PubMedGoogle Scholar
  27. Ohtsubo M, Theodoras AM, Schumacher J, Roberts JM, Pagano M (1995) Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol Cell Biol 15:2612–2624PubMedGoogle Scholar
  28. Rana SS, Bhasin DK, Nanda M, Singh K (2007) Parasitic infestations of the biliary tract. Curr Gastroenterol Rep 9:156–164PubMedCrossRefGoogle Scholar
  29. Rhoads ML, Fetterer RH (1997) Extracellular matrix: a tool for defining the extracorporeal function of parasite proteases. Parasitol Today 13:119–122PubMedCrossRefGoogle Scholar
  30. Rogoff HA, Kowalik TF (2004) Life, death and E2F: linking proliferation control and DNA damage signaling via E2F1. Cell Cycle 3:845–846PubMedGoogle Scholar
  31. Rosin MP, Saad DZS, Ward AJ, Anwar WA (1994) Involvement of inflammatory reactions and elevated cell proliferation in the development of bladder cancer in schistosomiasis patients. Mutat Res 305:283–292PubMedGoogle Scholar
  32. Shackelford RE, Kaufmann WK, Paules RS (1999) Cell cycle control, checkpoint mechanisms, and genotoxic stress. Environ Health Perspect 107(Suppl. 1):5–24PubMedCrossRefGoogle Scholar
  33. Slansky JE, Farnham PJ (1996) Transcriptional regulation of the dihydrofolate reductase gene. Bioessays 18:55–62PubMedCrossRefGoogle Scholar
  34. Sowers R, Toguchida J, Qin J, Meyers PA, Healey JH, Huvos A, Banerjee D, Bertino JR, Gorlick R (2003) mRNA expression levels of E2F transcription factors correlated with dihydrofolate reductase, reduced folate carrier, and thymidylate synthase mRNA expression in osteosarcoma. Mol Cancer Ther 2:535–541PubMedGoogle Scholar
  35. Thuwajit C, Thuwajit P, Kaewkes S, Sripa B, Uchida K, Miwa M, Wongkham S (2004) Increased cell proliferation of mouse fibroblast NIH-3T3 in vitro induced by excretory/secretory product(s) from Opisthorchis viverrini. Parasitology 129:455–464PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Young Ju Kim
    • 1
  • Min-Ho Choi
    • 1
  • Sung-Tae Hong
    • 1
  • Young Mee Bae
    • 1
  1. 1.Department of Parasitology and Tropical MedicineSeoul National University College of MedicineSeoulSouth Korea

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