OBJECTIVE: To explore clinical evaluate of CK19 mRNA and EGFR mRNA for diagnosis of laryngeal carcinoma micrometastasis, correlation between circulation tumor cell and lymph node metastasis. METHOD: Of 30 nude mice, 25 were randomly divided into 5 experimental groups (5 mice in each group), 5 were acted as control group. The mice were killed 2,4,6,8 and 10 weeks after injection. The expression of CK19 and EGFR mRNA in the peripheral blood and tumor tissue were detected by RT-PCR assay. The expression of EGFR in tumor tissue were detected by immunohistochemistry and lymph node transfer were confirmed using continuous pathological dying. RESULT: None of CK19 and EGFR mRNA were detected in peripheral blood of control group, CK19 mRNA-positive rate was 48% and 80% in peripheral blood and tumor tissue from the experimental group, respectively, and EGFR mRNA-positive rate was 36% and 76%, respectively. Lymph node metastasis happened in the exponential growth phase and transfer rate was 60%(15/25). The expression of CK19 mRNA and EGFR mRNA in lymphatic metastasis groups was higher than that of control, with a positive correlation between lymphatic metastasis and CTC (r = 0.655 , P < 0.01). The protein positive expression rate of EGFR were 88%(22/25) in tumor tissue. All peripheral blood expressed EGFR concomitant EGFR expressing in tumor tissues, and a high expression of EGFR in tumor tissue displayed high expression of EGFR in peripheral blood as well. CONCLUSION The expression of CK19 and EGFR mRNA in the peripheral blood can provide predictive information of lymphatic metastasis, EGFR mRNA might be a new target of treatment and diagnosis for malignant tumour.
Animals ; Disease Models, Animal ; ErbB Receptors ; blood ; Keratin-19 ; blood ; Laryngeal Neoplasms ; blood ; pathology ; Lymphatic Metastasis ; Mice ; Mice, Nude ; Neoplasm Staging ; RNA, Messenger ; genetics

BACKGROUNDAs the clinical value of cytokeratin-19 (CK19) and thymidine kinase-1 (TK1) in advanced gastrointestinal cancer remains controversial, we investigated their expression and clinical significance in this disease.

METHODSA total of 171 advanced gastrointestinal cancer patients were prospectively enrolled in this study. The mRNA level of CK19 was detected using quantitative real-time reverse transcription-polymerase chain reaction (PCR) in all patients, along with a control group of fifty healthy individuals. Furthermore, detection of TK1 protein was carried out in 96 patients using a chemiluminescence dot blot assay. The primary endpoint was overall survival (OS) time.

RESULTSPositive CK19 mRNA expression was detected in 74 (43.3%) of the 171 patients and positive TK1 expression was detected in 66 (68.8%) of the 96 patients. Furthermore, of the 96 patients, 36 (37.5%) were positive for both TK1 protein and CK19 mRNA, 30 (31.3%) were negative for TK1 protein, and 15 (15.6%) were negative for TK1 protein and positive for CK19 mRNA. The results indicated that patients who were positive for CK19 mRNA expression had significantly shorter OS times than those who were negative for it (median OS 7.7 vs. 9.7 months, respectively; P = 0.02). Moreover, patients who were positive for CK19 mRNA and TK1 protein expression had shorter OS times (median OS 6.1 months) than those who were positive for CK19 mRNA and negative for TK1 protein expression (median OS 9.1 months; P = 0.028). Positive CK19 mRNA expression was significantly associated with shorter OS in the univariate analysis (P = 0.027). Based on a multivariate Cox regression analysis, CK19 mRNA together with TK1 protein expression (P = 0.024) was an independent predictor for OS in gastrointestinal cancer patients.

CONCLUSIONSOur results suggest that positive expression of CK19 mRNA and TK1 protein is closely correlated with poor prognosis in advanced gastrointestinal cancer. Furthermore, both CK19 and TK1 are possible gastrointestinal cancer biomarkers.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; genetics ; Female ; Gastrointestinal Neoplasms ; blood ; genetics ; mortality ; Humans ; Keratin-19 ; blood ; genetics ; Male ; Middle Aged ; Prospective Studies ; Thymidine Kinase ; blood ; genetics

PURPOSE: The roles of circulating tumor cells (CTCs) as predictive and prognostic factors, as well as key mediators in the metastatic cascade, have been investigated. This study aimed to validate a method to quantify CTCs in peripheral blood using a real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay for cytokeratin (CK)-19 and to evaluate the utility of this assay in detecting CTCs in breast cancer patients. MATERIALS AND METHODS: Real-time monitoring PCR of fluorescently labeled specific hybridization probes for CK-19 mRNA was established. Peripheral blood samples from 30 healthy donors, 69 patients with early breast cancer, 47 patients with locally advanced breast cancer, and 126 patients with metastatic breast cancer were prospectively obtained and analyzed for CTC detection. RESULTS: CK-19 mRNA was not detectable in healthy subjects using the real-time RT-PCR method. The detection rates of CK-19 mRNA in breast cancer patients were 47.8% for early breast cancer (33/69), 46.8% for locally advanced breast cancer (22/47), and 61.1% for metastatic breast cancer (77/129). The detection rate of CK-19-positive CTCs in metastatic disease was slightly higher than early or locally advanced breast cancer; however, the detection rate according to disease burden was not statistically different (p=0.097). The detection rate was higher in patients with pleural metastasis (p=0.045). CTC detection was associated with poor survival (p=0.014). CONCLUSION: A highly specific and sensitive CK-19 mRNA-based method to detect CTCs in peripheral blood in breast cancer patients can be used in further prospective studies to evaluate the predictive and prognostic importance of CTCs.
Biomarkers, Tumor/*blood ; Breast Neoplasms/blood/*pathology ; Female ; Humans ; Keratin-19/*blood/genetics ; *Neoplastic Cells, Circulating ; Prognosis ; Prospective Studies ; RNA, Messenger/*blood ; Real-Time Polymerase Chain Reaction ; *Reverse Transcriptase Polymerase Chain Reaction/methods

Adult ; Aged ; Biomarkers, Tumor ; analysis ; Carcinoma, Non-Small-Cell Lung ; blood ; genetics ; pathology ; Cell Differentiation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Keratin-19 ; genetics ; Leukocytes ; metabolism ; Lung Neoplasms ; blood ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Survival Analysis