Cellular and Molecular Biology

Tumor DNA circulating in the plasma might play a role in metastasis. The hypothesis of the genometastasis

D. García-Olmo1, D.C. García-Olmo1, J. Ontañón1, E. Martinez1 and M. Vallejo2

1Experimental Research Unit and Department of General Surgery, Albacete General Hospital, Albacete, Spain and
2Reproductive Endocrine Unit, Dept. of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

Offprint requests to: Dr. Damián García-Olmo, Unidad de Investigación, Hospital General de Albacete, Hnos. Falcó s/n. 02006 Albacete, Spain. FAX: 34-967 24 39 52. e-mail: dgolmo@arrakis.es


Summary. Background: Clinical and experimental observations suggest that more than one pathway might be involved in the development of metastases. In the present study, we examined the presence of tumor DNA in plasma using an experimental model in which tumor cells were modified with a genome-associated tag. We also investigated whether plasma of tumor-bearing rats had any effect on cultured cells and healthy animals. Methods: Transfected cancer cells (DHD/K12-PROb stably transfected with pCDNA3.1CAT.) were injected subcutaneously into the chest of BD-IX rats. Animals were divided into ten groups according to the time between injection of tumor cells and euthanasia. Prior to euthanasia (2-14 week), blood samples were collected by cardiac puncture. To detect circulating tumor cells and CAT-encoding DNA in plasma, we performed PCR with nested primers. Fifty samples of plasma were chosen at random to supplement the medium of fifty cultures of DHD cells for 10-12 days. PCR for the detection of CAT DNA in cells was performed approximately one to two months later. Four healthy rats received an intraperitoneal injection of plasma from a tumor-bearing rat five times at week for 4 to 6 weeks. Animals were sacrificed and samples of liver, kidney, spleen, omentum, blood and lung were processed by PCR for the detection of CAT DNA. Results: Detection of CAT DNA in plasma was slightly more frequent than in the buffy-coat fraction. All surviving cultures that had been supplemented with plasma were positive at some point for CAT DNA. In all four healthy animals injected with plasma of tumor-bearing rats, the marker gene for CAT was found in extracts of lungs. Conclusion: Our present observation lead us to propose the following hypothesis. Metastases might develop as a result of transfection of susceptible cells in distant target organs with dominant oncogenes that are present in the circulating plasma and are derived from the primary tumor. Histol. Histopathol. 14, 1159-1164 (1999)

Key words: Tumor DNA, Plasma, Metastasis pathways, Colon adenocarcinoma, Rat

DOI: 10.14670/HH-14.1159