Abstract
Nasopharyngeal carcinomas (NPC) are tumors of nasopharynx origin with high rate of distance metastases after radiotherapy. Therefore, detection of micrometastasis will be an important issue in the prognostic and the choice of the systemic treatment. Our aim was early detection of circulating tumor cells in the blood of NPC patients by an RT-PCR assay for cytokeratin19 mRNA. Patients and methods used are venous blood samples obtained from 30 NPC patients with biopsy-proven NPC and 20 healthy blood donors which were tested using a previously reported assay. Identity of products was confirmed by sequencing. Results obtained show that 10 of the 30 NPC were positive for CK19 transcripts in peripheral blood, suggesting the presence of tumour epithelial cells. No CK19 positive cells were detected in the 20 healthy volunteers. Nevertheless, we encountered the problem of CK19 pseudogene interference due to genomic DNA contamination of RNA preparations. Four of the 10 positive patients were proven to have CK19 mRNA, as assessed by sequencing. In the remaining six, the presence of the CK19 pseudogene may has masked the CK19 mRNA in RT-PCR and sequencing. In conclusion, we described an improved RT-PCR assay that is sensitive and has high clinical specificity to detect minimal metastatic disease in NPC patients. Nevertheless, one should be aware about the necessity to optimize the analytical specificity of the assay. Key words: Nasopharyngeal carcinomas, micrometastasis, cytokeratin 19, nested reverse transcriptase polymerase.
Highlights
Nasopharyngeal Carcinoma (NPC) is a rare tumor arising from the mucosal epithelium of the nasopharynx
We studied a cohort of 30 Tunisian patients with non apparent but biopsy-proven Nasopharyngeal carcinomas (NPC)
This highlights the importance to understand the molecular process of metastase and the necessity to establish a synoptic table of markers which allow easy prediction, detection and localization of active metastasis and identification and definition of therapeutic targets
Summary
Nasopharyngeal Carcinoma (NPC) is a rare tumor arising from the mucosal epithelium of the nasopharynx. It is vastly more common in South East Asia and North Africa than elsewhere (Chen et al, 2013). Patients present at the moment of diagnosis have occult or detectable metastases. NPC is difficult to detect early due to the anatomical position of the nasopharynx, its symptoms that mimic those of many other conditions and its tendency to present with cervical lymph node metastases. Early detection of micrometastases and the prediction of recurrence will be an important issue in the prognostic of newly diagnosed NPC patients, in the choice of systemic treatment and in monitoring the effectiveness of the adjuvant therapy
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