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

OBJECTIVE: To investigate the correlation of the nuclear factor (NF-kappaB) and laryngocarcinoma circulating tumor cells (CTC), observe nuclear factor inhibitor PDTC on laryngeal cancer CTC and its possible mechanism. METHOD: In order to establish of laryngocarcinoma nude mice model ,The nude mice inoculated with laryngo carcinoma Hep-2 cells. To 50 nude mice were randomly divided into experimental group (PDTC pretreatment group), the control group (saline group), and Sham group (not inoculated carcinoma cells), tumor inoculated mice were labeled with CK-19 CTC markers, and sacrificed after 8 weeks, then RT-PCR detect CK-19mRNA expression in peripheral blood as to understand the expression of laryngocarcinoma CTC; removed tumor to determine its size, immunohistochemical expression of NF-kappaB in laryngocarcinoma, real-time fluorescence Quantitative PCR detection of laryngocarcinoma VEGF and MMP-9mRNA expression. RESULT: CTC occurrence and the expression of NF-kappaB was not obviously associated, expression in the CTC-positive laryngeal carcinoma, NF-kappaB-positive rate was 90%, while the expression of negative CTC laryngocarcinoma, NF-kappaB-positive rate was 56.7% (P>0.05), but connected with the activity of NF-kappaB; PDTC can inhibit NF-kappaB activation, reduce the incidence of CTC in laryngocarcinoma nude mice model, PDTC group CTC positive rate of 5%, significantly lower than the control group CTC positive rate of 45% (P<0.01). CONCLUSION PDTC can reduce the incidence of CTC in the laryngocarcinoma nude mouse model, its role may benefit from PDTC reduced the ability of laryngocarcinoma metastasis, which may be as PDTC inhibit NF-kappaB activity, thus make VEGF-C and the expression of MMP-9 down, reducing angiogenesis and reduce tumor invasion of the larynx.
Animals ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; NF-kappa B ; metabolism ; Neoplastic Cells, Circulating ; Proline ; analogs & derivatives ; pharmacology ; Thiocarbamates ; pharmacology ; Vascular Endothelial Growth Factor C ; metabolism

OBJECTIVE: To investigate the expression of glucose transporter 1 (GLUT1) in human laryngeal squamous cell carcinoma and its relationship to clinical pathologic factors. METHOD: Tumor tissues and the peritumoral and normal laryngeal tissue were collected from 57 patients with laryngeal squamous cell carcinoma. The expression of GLUT1 was evaluated at protein and mRNA level by immunohistochemistry and reverse transcription polymerase chain reaction(RT-PCR) in relation to clinical pathologic characteristics. RESULT: GLUT1 protein and mRNA expression were significantly higher in tumoral tissues than in peritumoral and normal tissues (P < 0.01, respectively). Poorly differentiated tumor expressed higher levels of GLUT1 protein and mRNA compared to well differentiated ones (P < 0.05, respectively). Stronger GLUT1 protein and mRNA were detected in larger tumors compared with that in smaller ones (P < 0.05, respectively). There were significant differences in GLUT1 protein and mRNA expression in relation to TNM stage (P < 0.05, respectively). The expression levels of GLUT1 protein and mRNA were positively correlated with lymphatic metastasis (P < 0.05, respectively). CONCLUSION GLUT1 may play an important role in tumorigenesis, development, differentiation, lymphatic metastasis and prognosis of human laryngeal squamous cell carcinoma, and may be a useful marker of diagnosis and prognosis.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Glucose Transporter Type 1 ; metabolism ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; RNA, Messenger ; genetics