Increased cell proliferation of mouse fibroblast NIH-3T3 in vitro induced by excretory/secretory product(s) from Opisthorchis viverrini

C. THUWAJIT a1a4c1, P. THUWAJIT a1a4, S. KAEWKES a2a4, B. SRIPA a3a4, K. UCHIDA a5, M. MIWA a5 and S. WONGKHAM a1a4
a1 Department of Biochemistry, Khon Kaen University, Khon Kaen 40002, Thailand
a2 Department of Parasitology, Khon Kaen University, Khon Kaen 40002, Thailand
a3 Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
a4 Liver fluke and Cholangiocarcinoma Research Center, Khon Kaen University, Khon Kaen 40002, Thailand
a5 Department of Biochemistry and Molecular Oncology, Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan

Article author query
thuwajit c   [PubMed][Google Scholar] 
thuwajit p   [PubMed][Google Scholar] 
kaewkes s   [PubMed][Google Scholar] 
sripa b   [PubMed][Google Scholar] 
uchida k   [PubMed][Google Scholar] 
miwa m   [PubMed][Google Scholar] 
wongkham s   [PubMed][Google Scholar] 


Infection by Opisthorchis viverrini is a strong risk factor for cholangiocarcinoma. However, the mechanism by which the parasite is involved in carcinogenesis is not clear. In addition to the direct damage of the bile duct epithelium via direct contact with O. viverrini, the excretory/secretory (ES) product(s) released from the parasites may play important roles in this process. We therefore investigated the responses of a fibroblast cell line, NIH-3T3, to ES product(s) released from O. viverrini by using a non-contact co-culture technique. In this culture system, the parasites in the upper chamber had no direct contact with the NIH-3T3 cells in the lower chamber of the culture plate. The results indicated a marked increase in NIH-3T3 cell proliferation in the non-contact co-culture condition with either 0% or 10% calf serum in the medium compared with that without parasites. ES product(s) increased cell proliferation by stimulating the expression of phosphorylated retinoblastoma (pRB) and cyclin D1, the key proteins in driving cells through the G1/S transition point of the cell cycle. This led to the induction of cells going into the S-phase of the cell cycle. ES product(s) also changed the morphology of NIH-3T3 cells to a refractive and narrow shape, which allowed the cells to proliferate in the limited culture area. For the first time, we have been able to demonstrate increased cell proliferation induced by the ES product(s) from O. viverrini; this finding may clarify how O. viverrini ES product(s) affect human bile duct epithelium during cholangiocarcinogenesis.

(Received January 14 2004)
(Revised March 4 2004)
(Revised March 11 2004)
(Accepted March 12 2004)

Key Words: Opisthorchis viverrini; excretory/secretory product(s); NIH-3T3; non-contact co-culture; cell proliferation; cell cycle.

c1 Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Tel: +66-43-348386. Fax: +66-43-348386. E-mail: