Expression Of Hypoxia Induced Factor-1α, Glucose Transporter Protein-1 In Cervical Squamous Cancer, And Correlation With Angiogenesis

Expression of Hypoxia Induced Factor-1α, Glucose Transporter Protein-1 in Cervial Squamous Cancer, and Correlation with AngiogenesisObjectiveCervical cancer is the most common gynecological carcinoma, which seriously threatens the health of women. Due to the wide application of cervical cytology filtering and the advancement of HPV detecting technology, prevention,earlier discovery and therapy of cervical cancer have been enabled in recent 40 years. Regarding cervical cancer, two topics, i. e., how to control cervical cancer and the mechanism on cancer metastasis, emerged out as currently most important issues.The hypoxia-inducible factor lα(HIF-1α) is able to combine with specific region points of hypoxia response element (HRE), and then initiate the induction of target gene, and therefore the organism can generate a series of adaptive responses to hypoxia. Glucose transporter-1 (GLUT-1) is an important target gene of HIF-1α. These two factors are highly involved with the generation and evolution of tumor, and therefore they become more popular in recent research. Yet, there is few research on the correlation between their expressions in cervical cancer. This paper discusses the expression in different cervical tissues using immunohistochemistry, and confirms the correlation between these two factors. In addition, it also discusses the relation between these two factors and the generation of tumor vessels.Materials and Method1. Objects34 paraffin samples of cervical cancer without radiotherapy or chemotherapy, were collected from China Medical University affiliated Shen Jing Hospital, January, 2002 to December, 2005. The ages of inpatients range from 25 to 70, and the median age is 43. According to FIGO2000 clinical stage standard, 16 of the 34 samples belong to disease stage 1, while the rest 18 distribute in stage 2.15 samples are with a greater tumor than 4cm, while the rest 19 are with one less than 4cm. 13 samples belong to high differentiation, while the rest 21 samples belong to medium or low differentiation. 19 samples are with pelvic lympt mode metastasis status, while the rest 15 samples are not. In addition, 10 CIN samples and 10 normal cervical tissue samples are corrected for comparison.2. ReagentCondensed rabbit anti-human HIF-1αpolyclonal antibody, was purchased from Wuhan Boshide biology Ltd. Corp. ready-to-use rabbit anti-human GLUT-1 monoclonal antibody, ready-to-use rabbit anti-human CD34 monoclonal antibody, streptavidin peroxidase (SP) hypersensitivity reagent cases and diaminobenzidine (DAB), were purchased from Fuzhou Maixin Biology Ltd. Corp. Self confected citric acid antigen repaired buffered liquid and 3% H₂O₂ liquid.3. Experiment MethodThe experiment was conducted using immunohistochemistry, and according to instructions of the reagent cases. The first antibody dilution concentration is 1: 200. This research used pre-existing breast cancer masculine sections as comparison samples, and used phosphate buffered saline (PBS) instead of the first antibody as lank comparison samples. Every sample is stained with Hematoxylin-eosin.3. Results Determination(1) HIF-1αand GLUT-1According to the method of Batchtiary et al., a semi-quantitive evaluation mechanism is processed, which combines the intensity and the percentage of masculine cells out of total cells. Exactly, a poor transfected sample is assigned with 1 point, a clear transfected sample is assigned with 2 points, a strong transfected sample is assigned with 3 points. If the percentage of masculine cells lies from 0% to 10%, it is assigned with 1 point; if from 11% to 50%, it is assigned with 2 points; if from 51% to 80%, it is assigned with 3 points, if over 80%, it is assigned with 4 points. The final evaluation is calculated as the total of the above two scores. If the total is less than 3, it is determined as negative expression (-); if greater than 3, it is positive expression (+), if greater than 5, it is medium positive expression (++); if greater than 7, it is strong positive expression (+++).(2) MVD quantitive evaluationAccording to the Weidner's method, CD34 monoclone antibody is used to mark vessel endothelium cells. An individual micro vessel is represented by cancer stromal isolated buffy vascular endothelium cell, cell cluster or where the vessel cavity is less than 8 erythrocytes but has no muscular layer on vascular wall. The max MVD inside tumor tissues is first determined under low zoom views, then count MVD of 3 views under high zoom view and calculate average.3. Statistical AnalysisSPSS 13.0 software is used for statistics calculation. The comparison of rate is performed usingχ² test or Fisher's exact possibility test of crosstabs. Grade correlation is analyzed with Spearman method. Probability of P<0.05 was considered statistically significant.Resultsâ‘ The expression rates of HIF-1αand GLUT-1 in cervical cancer and CIN (Cervical Intraepithelial Neoplasia) were significantly higher than those in normal cervical tissue and CIN.â‘¡In cervical cancer, there was no relationship between HIF-1αexpression and age, tumor size, pathologic type or histological differentiation, but associated with disease stage and pelvic lympt mode metastasis status; the expression of GLUT-1 was related with histological differentiation, pelvic lymph mode metastasis and disease stage, but not with age, tumor size and pathologic type.â‘¢There were positive correlations between the expression of HIF-1α, GLUT-1 and MVD.Conclusionâ‘ The over-expression of HIF-1αand GLUT-1 may be associated with the carcinogenesis and development of cervical cancer, and they therefore may both contribute to neoangiogenesis.â‘¡The expression of GLUT-1 may be enhanced by HIF-1α.