Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100)

AM: To establish and characterize a new cholangiocarcinoma cell line from a patient living in the Opisthorchis viverrini (O. viverrini) endemic area of Northeast Thailand. METHODS: Fresh liver biopsy and bile specimens were obtained from a 65-year-old Thai woman with cholangiocarcinoma of the ports hepatis. After digestion, the cells were cultured in Ham's F12 media. The established cell line was then characterized for growth kinetics, cell morphology, imm-unocytochemistry and cytogenetics. Tumorigenicity of the cell line was determined by heterotransplanting in nude mice. RESULTS: The primary tumor was a poorly differentiated tubular adenocarcinoma. Examination of the bile revealed malignant cells with O. viverrin eggs. The cholangiocarcinoma cell line KKU-100 was established 4 mo after the primary culture-population doubling time was 72 h. KKU-100 possesses compact and polygonal-shaped epithelial cells. Immunocytochemically, this cell line exhibited cytokeratin, EMA, CEA, and CA125, but not a-fetoprotein (AFP), CA19-9, desmin, c-met, or p53. Such protein expressions parallel those of the primary tumor. Cytogenetic analysis identified aneuploidy karyotypes with a modal chromosome number of 78 and marked chromosomal structural changes. Inoculation of KKU-100 cells into nude mice produced a transplantable, poorly differentiated aden-ocarcinoma, similar to the original tumor. CONCLUSION: KKU-100 is the first egg-proven, Opisthorchis- associated cholangiocarcinoma cell line, which should prove useful for further investigations of the tumor biology of this cancer.

Opisthorchiasis in Thailand: Review and current status

Opisthorchiasis caused by Opisthorchis viverrini （Oviverrini） remains a major public health problem in many parts of Southeast Asia including Thailand, Lao PDR, Vietnam and Cambodia. The infection is associated with a number of hepatobUiary diseases, including cholangitis, obstructive jaundice, hepatomegaly, cholecystitis, cholelithiasis and cholangiocarcinoma. The liver fluke infection was induced by eating raw or uncooked fish products that is the tradition and popular in the northeastern and northern region, particularly in rural areas of Thailand. Health education programs to prevent and control opisthorchiasis are still required in high-risk areas.

Opisthorchiasis-associated biliary stones:Light and scanning electron microscopic study

AIM:Biliary stones are frequently encountered in areas endemic for opisthorchiasis in Thailand.The present study was to describe the prevalence and pathogenesis of these stones. METHODS:Gallstones and/or common bile duct stones and bile specimens from 113 consecutive cholecystectomies were included.Bile samples,including sludge and/or microcalculi,were examined for Opisthorchis viverrinieggs, calcium and bilirubin.The stones were also processed for scanning electron microscopic(SEM)study. RESULTS:Of the 113 cases,82 had pigment stones,while one had cholesterol stones.The other 30 cases had no stones.Most of the stone cases(76%,63/83)had multiple stones,while the remainder had a single stone.Stones were more frequently observed in females.Bile examination was positive for O.viverrini eggs in 50% of the cases studied.Aggregates of calcium bilirubinate precipitates were observed in all cases with sludge.Deposition of calcium bilirubinate on the eggshell was visualized by special staining.A SEM study demonstrated the presence of the parasite eggs in the stones.Numerous crystals, morphologically consistent with calcium derivatives and cholesterol precipitates,were seen. CONCLUSION:Northeast Thailand has a high prevalence of pigment stones,as observed at the cholecystectomy,and liver fluke infestation seems involved in the pathogenesis of stone formation.

Parasites of the liver:A global problem?

Parasitic liver diseases can be caused by trematodes,cestodes,nematodes,and protozoa.This pathology is significant because millions of people in different parts of the world have liver parasites,which can manifest themselves in the development of inflammation,liver cysts,cholecystitis,cholelithiasis,pancreatitis and liver cirrhosis that are often threatening their lives.The International Agency for Research on Cancer considers three species of trematodes,Schistosoma haematobium,Opisthorchis viverrini and Clonorchis sinensis,to be carcinogens.Complex modern examination methods,in some cases including extensive screening of large populations,are required for diagnosing liver parasites.Treatment of parasitic liver diseases is differentiated and can involve a combination of surgical and therapeutic measures.There is no doubt that the clinical and epidemiological scale allows one to regard parasitic liver diseases as a global healthcare problem.

Fish sharing as a risk factor for Opisthorchis viverrini infection:evidence from two villages in north-eastern Thailand

Background:Foodborne trematodiasis(FBT)is a significant global health problem,with the liver flukes Opisthorchis viverrini,O.felineus,and Clonorchis sinensis contributing to half of the global burden of FBT.North-eastern Thailand where O.viverrini is endemic and un-cooked fish dishes remain an integral part of the food culture has the highest reported incidence of opisthorchiasis,including associated cholangiocarcinoma.Both food sharing and eating practices are potentially important factors in FTB,suggesting an important role for the social ecology of disease transmission in these rural communities.Methods:Two rural Thai-Lao villages that were part of a 12-village project in Northeastern Thailand were selected for detailed investigation of O.viverrini infection risk associated with sharing of raw fish dishes among households.The project included screening individuals for infection and cholangiocarcinoma,a household questionnaire,and offering treatment options for positive individuals.Social network mapping was used to construct raw fish dish-sharing networks and create a proxy variable capturing variability in the degree of food sharing(DFS),measured as the number of different households with which each household shared fish dishes.Measures of associations between DFS,O.viverrini infection,the frequency of raw fish consumption,and the number of raw fish dishes consumed were generated using binary logistic regression,proportional odds ordinal logistic regression,and Poisson regression.Results:The results showed that the probability that a household has members infected with O.viverrini increased by~7%(P<0.01)for each additional household included in its network.Moreover,the frequency and number of types of raw fish dishes consumed increased significantly as the DFS increased.Of the two villages,that with the highest infection prevalence(48%versus 34.6%)had significantly higher social connectivity overall(P<0.001).Conclusions:Our findings suggest that the social ecology of human settlements may be key to understanding the transmission dynamics of some FBT.In the case of O.viverrini in Thai-Lao communities,for which food sharing is a traditional practice supporting social cohesion,food sharing network mapping should be incorporated into community-based interventions.These should encourage fish dish preparation methods that minimize infection risk by targeting households with high DFS values.