Fluorescent Quantitative RT-PCR Detection Of Micrometastasis In Peripheral Blood Of Patients With Nasopharyngeal Carcinoma

Objective To explore the expression of CK14 mRNA and EGFR mRNA in peripheral blood of 87 patients with nasopharyngeal carcinoma(NPC) and evaluate its value for diagnosis of micrometastasis.Methods 87 patients with NPC underwent detailed history, careful physical examination and complete relevant imaging examination. TNM stage , clinical stage , degree of histologic differentiation and syndrome type of Traditional Chinese Medicine(TCM ) were determined in all the cases before therapy. The expression of CK14 mRNA and EGFR mRNA in peripheral blood were detected by fluorescent quantitative RT-PCR(FQ-PCR).The correlation between peripheral blood micrometastasis and tumor TNM stage, clinical stage, degree of histologic differentiation and syndrome type of TCM of the patients with NPC was analyzed.Results 1. The rate of positive expression of CK14 mRNA and EGFR mRNA in peripheral blood of 87 patients with NPC was 64.4% (56/87) and 58.6% (51/87) respectively, even the combined assay produced a value of 71.3% (62/87) , meanwhile that in peripheral blood of 5 patients with distant metastasis and 5 tumor tissues were both 100%, furthermore none of 10 healthy individuals had positive expression in peripheral blood. 2. The rate of positive expression of CK14 mRNA and EGFR mRNA in peripheral blood of patients with NPC were not related to the age and gender (P>0.05), but were related to degree of histologic differentiation and clinical stage (P<0.05). 3. The level of expression of CK14 mRNA and EGFR mRNA in peripheral blood of patients with NPC were related to T stage, N stage and clinical stage(P<0.05), and the later stage of carcinoma, the higher level of expression. 4. The level of expression of CK14 mRNA and EGFR mRNA in peripheral blood of patients with NPC were related to degree of histologic differentiation (P<0.05), and the level of expression of poorly differentiated squamous cell carcinoma was higher than well differentiated squamous cell carcinoma. 5. The distribution of syndrome type of TCM of the patients with NPC at different clinical stages were statistically significant different (P<0.05), coagulation of qi and blood syndromes, accumulation of phlegm syndromes, stagnation of virulent fire syndromes and asthenia of healthy energy syndromes were mainly in I stage patients,Ⅱstage patients,Ⅲstage patients andⅣstage patients, even accounting for 54.5%(6/11),64% (16/25),61.5%(24/39), and 66.7%(8/12) respectively. Furthermore, the level of expression of CK14 mRNA and EGFR mRNA in peripheral blood of patients with NPC were related to syndrome type of TCM (P<0.05), asthenia of healthy energy syndromes> stagnation of virulent fire syndromes>accumulation of phlegm syndromes>coagulation of qi and blood syndromes.Conclusion 1. Detecting the expression of CK14 mRNA and EGFR mRNA in peripheral blood of patients with NPC by FQ-RCR can accurately reflect the situation of peripheral blood micrometastasis.2. There is correlation between peripheral blood micrometastasis and malignant degree and disease progression.3. The syndrome type of TCM of patients with NPC is closely related to clinical stage, with the progress of clinical stage, the syndrome type of TCM present its trend of general evolution"coagulation of qi and blood syndromes→accumulation of phlegm syndromes→stagnation of virulent fire syndromes→asthenia of healthy energy syndromes", and with the evolution of syndromes, the level of expression of CK14 mRNA and EGFR mRNA in peripheral blood gradually raise up.