Dissertation
Dissertation > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Uterine tumors

Detection of Terc,ezrin and Galectin-3 Clinical Pathological Significance in Cervical Squamous Cell Carcinoma by Manual Tissue Chip

Author YuanYanLong
Tutor HeChunNian
School Hebei Medical University
Course Pathology and Pathophysiology
Keywords Cervical squamous cell carcinoma Cervical intraepithelial neoplasia TERC gene Ezrin Galectin-3 Fluorescence in situ hybridization Immunohistochemistry Tissue microarray
CLC R737.33
Type Master's thesis
Year 2010
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Objective: Cervical cancer is one of the common malignant tumors that affect women's lives and health. Mortality after breast cancer, in second place. In recent years, with the overall conduct of the cervical cancer screening, the incidence rate has greatly reduced, but people never stopped research on early diagnosis and prevention and treatment of cervical cancer. Recent studies show that almost all cervical squamous cell dysplasia to cervical squamous cell carcinoma of the process of transforming the multiple copies of the long arm of chromosome, most important of which is located at 3q26.3 TERC gene, the gene expansion increasing block apoptosis, leading to tumorigenesis. Multiple copies of the long arm of chromosome 3 and TERC gene amplification in more than 90% of high-grade precancerous lesions and low-grade precancerous lesions of the sensitivity and specificity for identification of cervical cells, detection of TERC amplification most helpful cervical screening and early diagnosis of squamous cell carcinoma. Ezrin protein is a membrane-cytoskeleton junction proteins play an important role in the number of cells formed in the cell morphology, movement, adhesion and cell signal transduction activity. The various membrane extracellular signal integration Ezrin protein and membrane protein directly connected to the incoming cells, causing the tumor, its overexpression may play an important role in the occurrence and development of cervical squamous cell carcinoma. Its targeted therapy may become a new way of cancer treatment. Galectins-3 is the aggregation of a prime family involved in cell proliferation, differentiation, local immune regulation and apoptosis, tumor growth, adhesion, invasion and metastasis. Their research has the potential to provide new ideas and methods for the early diagnosis and treatment of cervical squamous cell carcinoma. The subject using tissue microarray, fluorescence in situ hybridization study the TERC gene, and immunohistochemical detection Ezrin, Galectin-3 protein in normal cervical squamous epithelium, CIN and cervical squamous cell carcinoma tissues explore TERC, Ezrin Galectin-3 with varying degrees of cervical lesions and clinical pathology in the three may exist between cervical squamous cell carcinoma role in the occurrence and development of their mutual relations, analyze its role in the diagnosis of cervical lesions. Method: a collection of cervical tissue samples of 196 cases, including 48 cases of cervical squamous cell carcinoma, cervical CIN Organization (120 cases, 40 cases in each grade CIN lesions), normal cervical tissues 28 patients. Applications homemade tissue microarray technology, 196 cases of donor specimens array design order turn into a recipient paraffin block, fusion, made of sliced ??need tissue microarray. 3 using fluorescence in situ hybridization detection of TERC amplification in cervical squamous cell carcinoma, cervical CIN organizations and normal cervical squamous epithelial tissue. 4 detection of cervical squamous cell carcinoma by immunohistochemistry (SP), the cervical CIN organizations, normal cervical squamous epithelium Ezrin and Galectin-3 protein expression. SPSS16.0 statistical software for statistical analysis, to alpha = 0.05 significance level, statistically significant when P LT; 0.05. Results: the design of a tissue microarray preparation and staining results based on topics and test methods require making a total of 7 × 7 array tissue microarray 4, organizational dot matrix arranged in neat rows, dot matrix shift phenomenon, three sites missing, 18-bit point no meaningful organization, other bits point organizations see a meaningful organization results total was meaningful sample of 175 cases (cervical squamous cell carcinoma 48 cases, 33 cases of CIN III, CIN Ⅱ 36 cases of, CIN Ⅰ 34 cases, 24 cases of normal cervical squamous epithelium) , the morphology of the tissue sites can be observed rate of 89.3%. HE staining, no off-chip, ectopic and wrinkles, fluorescence in situ hybridization signal is clear, clean background, immunohistochemical staining positive positioning clear, clean background, no off-chip phenomenon. 2 TERC gene in cervical epithelial amplified situation in the various lesions using fluorescence in situ hybridization method detected 48 cases of cervical squamous cell carcinoma, 103 cases of CIN (CIN III in 33 cases, 36 cases of CIN Ⅱ, CIN Ⅰ 34 cases of) including 24 cases of normal cervical squamous epithelium TERC gene amplification. The results showed: normal cervical squamous epithelium TERC gene amplification, CIN I and 11.8% (4/34), CIN II-44.4% (16/36), 83.3% of CIN III, 69.7% (23/33) and cervical squamous cell carcinoma (40/48), with the higher level of the lesion, the TERC gene amplification rate is gradually increased, the difference was statistically significant (p lt; 0.05); high-grade CIN lesions in multiple copies of chromosome 3 cells, cervical squamous cell carcinoma The detected significant amplification of the TERC gene and a lot of multiple copies of chromosome 3 in tumor cells. TERC gene in cervical squamous cell carcinoma, the amplification rate of CIN III and CIN Ⅱ was significantly higher than in CIN I and normal cervical squamous epithelium, the difference was statistically significant (p lt; 0.05); significant difference between CIN II and cervical squamous cell carcinoma (p lt; 0.05); CIN III and cervical squamous cell carcinoma, the difference was not statistically significant (p gt; 0.05) between the normal cervical squamous epithelium and CIN I. TERC gene amplification in the identification of high-grade CIN lesions (CIN III, CIN Ⅱ) with sensitivity in the low-grade CIN lesions (CIN I) was 56.5%, specificity 88.2%, accuracy 67%, positive predictive value of 90.7%, The negative predictive value of 50.0%. 3 Ezrin protein expression in cervical intraepithelial lesions by immunohistochemistry SP method detected 48 cases of cervical squamous cell carcinoma, 103 cases of CIN (including 33 cases of CIN Ⅲ, 36 cases of CIN Ⅱ, 34 cases of CIN I) and 24 cases of normal cervical squamous Ezrin protein expression in the epithelial tissue. Normal cervical squamous epithelium, Ezrin protein expression in the cell membrane, a small amount or no expression from normal cervical squamous epithelium 8.3% (2/24), CIN I and 26.5% (9/34), CIN II 58.3% (21 / 36), CIN III and 51.5% (17/33) to cervical squamous cell carcinoma, 66.7% (32/48), the strong positive expression rate was gradually increased and cytoplasmic expression, the difference was statistically significant (p <; 0.05) . Ezrin protein in cervical squamous cell carcinoma and CIN III, CIN II in strongly positive expression rate of higher than normal cervical squamous epithelium and CIN Ⅰ group, the difference was statistically significant (p lt; 0.05); cervical squamous cell carcinoma difference between CIN III, CIN Ⅱ was not statistically significant (p gt; 0.05); was no significant difference between the normal cervical squamous epithelium and CIN I (p GT; 0.05). 4 Galectin-3 in cervical epithelial expression of the situation in the various lesions using chemical SP Act of immunohistochemical detection of 48 cases of cervical squamous cell carcinoma, 103 cases of CIN (including 33 cases of CIN Ⅲ 36 cases of CIN Ⅱ, 34 cases of CIN Ⅰ) and 24 cases of normal cervical scales like epithelial tissue expression. Galectin-3 of the normal cervical squamous little expression in the cytoplasm of cells in the middle, from the normal cervical squamous epithelium 0% (0/24), CIN I 5.9% (2/34), CIN II 19.4% (7 / 36), CIN III and 24.2% (8/33) to 25.0% of cervical squamous cell carcinoma (12/48), the strong positive expression rate gradually increased and the expression of a small amount of the nucleus, the difference was statistically significant (p lt; 0.05 ). Galectin-3 in cervical squamous cell carcinoma, CIN III and CIN Ⅱ expression was significantly higher than that of normal cervical squamous epithelium, the difference was statistically significant (p lt; 0.05); cervical squamous cell carcinoma and CIN Ⅰ difference was statistically significant (p < ; 0.05); difference between the other groups were not statistically significant (p gt; 0.05). 5 cervical squamous cell carcinoma TERC gene amplification and Ezrin, Galectin-3 expression in cervical squamous cell TERC gene amplification and expression of Ezrin protein correlation (p lt; 0.05), both were positively Relevance, r = 0.516. TERC gene amplification, no correlation between the expression of Ezrin protein expression of Galectin-3 (p GT; 0.05) rTERC with galectin-3 = -0.052, r Ezrin and galectin-3 = -0.252. 6 cervical squamous TERC gene amplification in cancer tissues, Ezrin protein and Galectin-3 expression and patient age, tumor size, degree of differentiation, clinical stage, lymph node metastasis and pathological grading the relationship between the TERC gene amplification with age, sex, tumor There was no significant correlation (p GT size, lymph node metastasis and pathological grade; 0.05), a significant correlation (p lt; 0.05) with clinical stage. Ezrin expression and patient age, gender, tumor size, pathological grade and clinical stage had no significant correlation (p gt; 0.05), with lymph node metastasis was significantly correlated (p lt; 0.05). Galectin-3 expression with age, gender, tumor size, histological grade, clinical stage and lymph node metastasis had no significant correlation (p GT; 0.05). Conclusion: a fluorescence in situ hybridization detection of TERC amplification amplification in normal cervical tissue, significantly amplified in a high level of CIN lesions and cervical squamous cell carcinoma associated with chromosome 3 multiple copies of an increase in the number of cells in the The differential diagnosis of CIN I and CIN II clinical practical significance. Fluorescence in situ hybridization detection of TERC amplification higher specificity, positive predictive value, and accuracy in the identification of high-grade CIN lesions and low-grade CIN lesions clinically predictive value for the development of CIN lesions. 3 Immunohistochemistry was used to detect the positive expression rate of Ezrin protein from normal cervical squamous CIN to cervical squamous cell carcinoma increased gradually, may aid the diagnosis of cervical lesions with clinical value. 4 Immunohistochemistry was used to detect the positive expression rate of Galectin-3 from normal cervical squamous CIN to cervical squamous cell carcinoma was gradually increased. 5 cervical squamous cell carcinoma, abnormal TERC gene amplification and Ezrin protein expression correlation was positive correlation between. Tips TERC gene and Ezrin protein collaborative detection in cervical squamous cell carcinoma of the diagnostic process. 6 in cervical squamous cell carcinoma of the TERC gene amplification with clinical stage correlation Ezrin protein abnormalities correlated with lymph node metastasis correlation. The 7 tissue microarray hand-made, the program is simple, economical and convenient, is a simple and feasible, efficient and stable technology, to meet the needs of related research projects and practical work.

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