Clinical impact of circulating tumor cells (CTC) morphological and molecular (ß catenin mutations) characterization in patients with hepatocellular carcinoma

G. Vona1, L. Estepa2, D. Damotte3, F. Capron4, C Béroud2, H Fontaine5, A. Mineur5, B. Nalpas5, H. Zylberberg5, D. Franco6, B. Lacour2, S. Pol5, C. Bréchot1,5, P. Paterlini-Bréchot1,2

1INSERM U370; Services 2Biochimie, 5Hepatologie, 3Anatomie-Pathologique, Hôpital Necker, Paris. 6Chirurgie Digestive, 4Anatomie-Pathologique, Hôpital A. Beclère, Clamart.

AIMS: The aim of this study was to detect and characterize CTC in patients with HCC by using ISET (Isolation by Size of Epithelial Tumor cells) (Vona, 2000). METHODS: Patients: 44 with HCC: 22 with localized HCC undergoing liver tumor resection, tested before and during surgery (group A), and 22 with diffused HCC, ineligible for surgery (group B), 30 with chronic active hepatitis (CAH), 42 with liver cirrhosis, 38 healthy individuals. Methods: ISET (5 ml of blood), microdissection of 60 single CTC (from 10 patients of group B), sequencing of ß-catenin exon 3. RESULTS: We failed to detect CTC and circulating tumor micremboli (CTM) in 38 healthy subjects, 30 patients with CAH and 42 patients with cirrhosis. CTC were found in 23/44 (5/22 from group A, tested before surgery, and 18/22 from group B) and CTM in 4/44 (1 from group A and 3 from group B) patients with HCC. In 9 patients from group A, CTC were absent before and appeared during surgery. The follow up study and actuarial curve analysis showed a significant correlation of the presence of CTC and/or CTM with a shorter survival (p=0.01). ß-catenin mutations were found in the reference cell line HuH6, at the expected position (positive control), and in 3/60 microdissected single CTC. CONCLUSIONS: Our results show that 1) this approach is feasible and specific for CTC and CTM detection 2) healthy subjects and patients with chronic liver disease scored negative 3) patients with diffused HCC have a higher number of CTC than patients with localized tumor 4) The presence and number of CTC significantly correlate with a shorter survival 5) ISET also allows detection of ß-catenin mutations in individual CTC and CTM.

For more information, contact

Paper presented at the International Symposium on Predictive Oncology and Intervention Strategies; Paris, France; February 9 - 12, 2002; in the section on Predictive Markers.

This presentation received an honorable mention in our poster contest and was recognized with the Symposium Presidents' Award for Scientific Excellence.