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Expression of Eag1in Rat Oral Mucosa Carcinogenesis

Author: SongPingPing
Tutor: NieMinHai
School: Luzhou Medical College
Course: Clinical Stomatology
Keywords: oral cancer Eagl Immunohistochemistry RT-PCR
CLC: R739.8
Type: Master's thesis
Year: 2012
Downloads: 10
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Abstract


objective:Oral squamous cell carcinoma (OSCC) is the most common oral cancer. It have high invasion and metastasis rate,however,due to its location and the special structure, such as the rich blood supply, more lymph node tissue, functional activities and other reasons。A large number of patients died each year because of this disease,there is poor quality of life and a low5-year survival rate in patients.Therefore it is very important to study of the diagnosis and treatment of oral cancer.OSCC mostly developed from oral precancerous lesion or precancerous condition. The early diagnosis and treatment are contributed to the prevention. An effective tumor marker has attracted much attention.It has been confirmed that the potassium channel KV10.1(Ether-a-go-go Eagl) highly expressed in many cancers and cancer cell lines, and thus infer that Eagl might be involved in the development of oral carcinoma.But the expression in oral cancer is unknown. Whether they have increased expression and site-specific,The correlationship with the cancer development process has to be conclusive.In view of this,We designed the experiment that is use4NQO (4-nitroquinoline-l-oxide) to induce oral carcinogenesis in SD rats, obtain the oral mucosa organization from each stage, such as the normal oral mucosa, precancerous lesions and oral squamous cell carcinoma.To detect the expession of protein and transription levels of Eagl gene at different time point.The relationship between Eagl and development of OSCC to be investigated,which would provide a new experimental base for prevention and treatment in OSCC.Methods:Fifty-eight male Sprague-Dawley (SD) rats weighing about200±10g,10to drink ordinary tap water as negative control and experiment group (48rats) was given drinking water containing40ppm4-NQO.All rats give basal diet until sacrificed. The forty-eight experimental rats were killed at the end of the9th,12th,18th and22nd weeks after the commencement of the experiment, respectively. The left ten controlled rats which were feed without4-NQO would be killed at the end of animal experiment. Extract the oral mucosa separately,one part fixed with4%neutral formaldehyde and another part stored in-70℃refrigerator from the same sample.Embedded with paraffin, cut into4μm thick serial sections and then pathological diagnosis of HE staining. Two experienced pathologists blinded method by HE staining for pathological diagnosis. SP immunohistochemical method was performed to detect Eagl protein expression and the expression levels were quantifyed;RT-PCR was performed to detect Eagl mRNA level,then the levels of Eagl protein and mRNA were statisticlly analyzed by SPSS17.0software package.Results:1.Get specimens of animal models including16cases of normal mucosa,11cases of mild dysplasia,12cases of moderate dysplasia, severe dysplasia10cases and9cases of squamous cell carcinoma.2.Immunohistochemistry result suggestted the expression of Eagl protein located in the cell memeberane and cytoplasm, no expression in the nucleus, no positive staining of normal mucosa. With the increase in the degree of abnormal proliferation of rat oral mucosa the Eagl protein expression was increased significantly. The Eagl positive constitute ratio is36.36%in the11cases of mild dysplasia organization;organization of12cases of moderate dysplasia tissues positive constitute ratio50.00%;10cases of severe dysplasia positive reach80%,9cases of oral squamous cell carcinoma, the positive ratio is88.89%.The Eagl expression in oral cancer rate significantly higher than those with mild to moderate dysplasia organization.The difference were statistically significant (P<0.05).3.The results of RT-PCR analysis showed that the Eagl mRNA level differences between the various pathological groups,there was statistically significant(P<0.05)。Conclusions:1.In the differentiation period organization of the4NQO-induced rat oral carcinogenesis process,the expression of Eagl protein with the cancer process was significantly increased.Imply the increase of Eagl expression along with the oral mucosa cancer occur in the early stage, its abnormal expression involved in the formation of the oral mucosa cancer. Transcriptional levels of Eagl in4NQO-induced rat oral carcinogenesis changes with statistical differences,and have the same trend with the protein expression.The enhancement of Eagl protein may be caused by abnomal gene transcription.3Eagl is attractive as target since this surface protein is virtually not detected in normal tissue outside the central nervous system, but is expressed in many tumors from different origins includ OSCC.

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CLC: > Medicine, health > Oncology > Oral cavity, maxillofacial tumors
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