Detection Of Terc,ezrin And Galectin-3 Clinical Pathological Significance In Cervical Squamous Cell Carcinoma By Manual Tissue Chip

Objective: Cervical cancer, one of the common malignant tumors, is harmful to women's lives and health.Deadth rate is second only breast cancer. In recent years, cervical cancer screening with fully operational, its incidence has been greatly reduced. Recent studies have shown that cervical cells from atypical hyperplasia to cervical cancer exist in the process of transforming chromosomal instability, almost all gain of 3q, most important of which is located at 3q26.3 of the TERC gene, TERC gene amplification can prevent cell apoptosis, and leading to tumorigenesis. The gain of 3q and TERC gene amplification in the differential diagnosis of cervical precancerous lesions and low-grade precancerous lesions of the sensitivity and specificity were 90%, and detection of TERC gene amplification contributed significantly to cervical cancer screening and early diagnosis. Ezrin is a membrane cytoskeletal connections protein. It play an important role in cell shape formation, movement, adhesion and cell signal transduction and many other cellular activities. A variety of external signals through Ezrin directly connected to the membrane integration of proteins incoming cells, and causing tumors. Its over-expression may be play an important role in the progression of cervical squamous cell carcinoma. Targeted therapy of Ezrin may become another new way of cancer treatment. Galectins-3 is a member of lectin family, participate in cell proliferation, differentiation, local immune regulation and apoptosis, and with relate to tumor growth, adhesion, invasion and metastasis. However, It is express in cervical lesions and its with clinical pathology relationship reports less and inconsistent. The research has provide new ideas and methods that early diagnosis and treatment of cervical cancer.This topic uses the tissue chip, Fluorescence In situ hybridization to research TERC, and uses the Immunohistochemistry to research the expression Ezrin and Galectin-3 protein in normal cervical squamous epithelia, CIN and cervical carcinoma.To analysis mutual relations of TERC, Ezrin and Galectin-3 abnormal expression in cancer progression. To investigate interrelations between TERC, Ezrin, Galectin-3 and clinical indicators of cervical cancer interrelationships.Methods:1 28 cases of normal cervical squamous epithelia, 120 cases of CIN and 48 cases of cervical carcinoma were collected.2 Put 196 cases cervical tissues by manual tissue chip technique into a recipient paraffin block, then fusing, slicing, gaining and baking,and made into tissue chip.3 Fluorescence In situ hybridization (FISH) was used to detect TERC gene amplification in cervical carcinoma, CIN and normal cervical squamous epithelia.4 Immunohistochemistry SP method was used to examine for the expression, relation and the effection of Ezrin and Galectin-3 in cervical carcinoma, CIN and normal cervical squamous epithelia.5 Analysis the data use spss16.0, significance was set at p<0.05.Results:1 The result of tissue chip4 tissue chips (7×7) were made. Dots of tissue chip lined up in order, uniformed in size and no shifting. 3 tissue dots were absence, 18 tissue no significant tissue structures. The significant tissue structures were watched in the other dots. The rate of complete tissue dots was 89.3%.The haematoxylin-eosin (HE) staining was uniformity and no dropping, moving and wrinkling.The immunohistochemical and FISH results show that the positive location was accurate and the background was clear.2 The Amplification of TERC geneIn normal cervical epithelia, the Amplification of TERC is absence,from CINⅠ11.8%(4/34), CINⅡ44.4%(16/36), CINⅢ69.7%(23/33) to SCC 83.3% (40/48), TERC gene amplification is significant differentiated at normal cervical squamous epithelia with CINⅠ, CINⅡ, CINⅢand cervical invasive squamous cell carcinoma (P<0.05). TERC gene amplification is significant differentiated at normal cervical epithelia, CINⅠwith CINⅡ, CINⅢ, cervical invasive squamous cell carcinoma (p<0.05). TERC gene amplification at cervical invasive squamous cell carcinoma with CINⅡis significant different(p<0.05), TERC gene amplification at cervical invasive squamous cell carcinoma with CINⅢand normal cervical epithelia with CINⅠno significant different(p>0.05), the sensitivity of TERC gene amplification was 56.5%, the specificity was 88.2%, the accuracy was 67.0%,the positive predictive value and negative predictive values were 90.7% and 50%.3 The expression of EzrinExpression of Ezrin in various lesions of cervix, 48 cases of cervical carcinoma, 103 cases of CIN and 24 cases of normal cervical seqamous epithelia were tested. From normal cervical epithelia 8.3%(29/24),CINⅠ26.5%(9/34),CINⅡ58.3%(21/36),CINⅢ51.5%(17/33) to SCC66.7%(32/48), there were significant differences among those groups (p<0.05). Ezrin expression is significant differentiated at normal cervical epithelia,CINⅠwith CINⅡ,CINⅢ,cervical invasive squamous cell carcinoma (p<0.05). Ezrin expression is no significant differentiated at normal cervical epithelia with CINⅠ(P>0.05),the expression of Ezrin at CINⅡ,CINⅢand SCC is no significant different too (P>0.05).4 The expression of Galectin-3Expression of Galectin-3 in various lesions of cervix, 48 cases of cervical carcinoma, 103 cases of CIN and 24 cases of normal cervical seqamous epithelia were tested. No expression of Galectin-3 was found in normal cervical epithela, the positive rates of Galectin-3 in CINⅠ,CINⅡ,CINⅢand cervical carcinoma group were 5.9%,19.4%,24.2%,25.0%, respectively. Galectin-3 expression is significant differentiated at normal cervical epithelia with CINⅡ, CINⅢand cervical invasive squamous cell carcinoma (p<0.05), and the expression at cervical invasive squamous cell carcinoma with CINⅠis significant different(p<0.05); other groups are no significant different(p> 0.05)。5 The relation of TERC,Ezrin,Galectin-3There was a positive correlation between amplification of TERC and expression of Ezrin(P<0.05). Between amplification of TERC and expression of Galectin-3,expression of Ezrin and Galectin-3 had no significant correlation.6 The relationship between Amplification of TERC, the expression of Ezrin,Galectin-3 and clinical pathological features of cervical carcinoma.No significant relationship were found between the amplification of TERC, the expression of Ezrin and clinical pathological features(age, sex, tumor size, pathological rank). However, there was significant relationship between the amplification of TERC and clinical stage, the expression of Ezrin and metastasis. No significant relationship were found between Galectin-3 and all clinical pathological features.Conclusions:1 There was no TERC gene amplification in normal cervical tissuse ,and was significant amplification in CINⅢand cervical cancinoma, and company by the 3 chromosome multiploid cell increase. There was signi -ficant clinical practical significance in antidiastole CINⅡand CINⅢ.2 Fluorescence in situ hybridization detection of TERC gene amplification to detect high-grade and low-grade CIN lesions have a higher specificity, positive predictive value and accuracy. There was significant clinical predictive significance in CIN.3 Ezrin overexpression rates become higher and higher from normal cervical squamous epithelia, CIN to cervical cancinoma.4 Galectin-3 overexpression rates become higher and higher from normal cervical squamous epithelia, CIN to cervical cancinoma.5 There was a positive correlation between amplification of TERC and the expression of Ezrin in occurrence and development of cervical squamous cell carcinoma. Probably signifient correlation of TERC and Ezrin in diagnose cervical squamous cell carcinomas. 6 There was significant relationship between the amplification of TERC and clinical stage. There was significant relationship between expression of Ezrin and metastasis. But with clinical pathological features was no significant relationship.7 Mannual tissue chip technique is simple, cost-effective, and reliable. This technique can provide a highly efficient, high-throughput mechanism for some research.