本文選題：膀胱癌 + 免疫組化��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:1、對前期獲得的cDNA全長序列行生物信息學(xué)研究,提交NCBI獲取基因號KR780251,蛋白號AKO62675。2、根據(jù)AKO62675與CK20表達(dá)差異性較大的序列設(shè)計多肽,制備AKO62675抗體:Anti-KR;3、根據(jù)AKO62675在膀胱癌組織、腺性膀胱炎以及正常膀胱組織中的相關(guān)表達(dá)情況,分析AKO62675與膀胱癌的發(fā)生發(fā)展是否具有相關(guān)性。方法:1、對cDNA全長序列進(jìn)行染色體定位、蛋白分析等,并利用sequin軟件向NCBI提交獲取基因號:KR780251以及蛋白號:AKO62675。2、根據(jù)計算機(jī)分析AKO62675與CK20表達(dá)差異性較大的序列,以此進(jìn)行多肽合成,免疫兔子并獲取多克隆抗體Anti-KR。3、采用免疫組化技術(shù)對40例的膀胱癌組織、15例的腺性膀胱炎組織以及23例的正常膀胱黏膜組織行AKO62675表達(dá)的檢測,對AKO62675的表達(dá)情況進(jìn)行了比較,了解該片段在膀胱腫瘤組織、腺性膀胱炎組織以及正常膀胱黏膜組織間的表達(dá)情況,分析該片段與膀胱癌的發(fā)生與發(fā)展是否有關(guān)系。結(jié)果:1、分析發(fā)現(xiàn)該序列與CK20在染色體上所處的具體位置不同,提交NCBI獲取基因號:KR780251以及蛋白號:AKO62675。2、應(yīng)用ELISA方法檢測成功制備出的多克隆抗體:Anti-KR,滴度1:64000,達(dá)到檢測要求。3、對膀胱癌組織、腺性膀胱炎及正常膀胱組織進(jìn)行免疫組化染色:在正常的膀胱黏膜中,Anti-KR染色陽性率達(dá)34.78%,而膀胱癌的組織中,Anti-KR染色陽性率達(dá)100.00%,在腺性膀胱炎組織中,Anti-KR染色陽性率也達(dá)100.00%,但染色較膀胱癌組織淺。結(jié)論:1、EST片段的全長序列KR780251有1839bp,含有完整的ORF,KR780251編碼的蛋白AKO62675可能是正常膀胱組織基因的突變體。2、AKO62675在膀胱癌中具有較高的表達(dá)性。
[Abstract]:Objective to study the full-length sequence of cDNA obtained in the early stage by bioinformatics, and to submit to NCBI the gene number KR780251, protein number AKO62675.2. According to the sequence of AKO62675 and CK20, we designed the peptide to prepare AKO62675 antibody: Anti-KRR3, and then prepared AKO62675 antibody: Anti-KR3 in bladder cancer tissue according to AKO62675. The expression of AKO62675 in cystitis glandularis and normal bladder tissues was analyzed. Methods: the cDNA full-length sequence was located on chromosome, protein was analyzed, and the gene number: KR780251 and protein number: AKO62675.2 were submitted to NCBI by sequin software. According to the computer analysis of the sequences in which the expression of AKO62675 and CK20 were different, the peptide was synthesized. Rabbits were immunized and polyclonal antibody Anti-KR.3was obtained. The expression of AKO62675 was detected by immunohistochemical technique in 15 cases of cystitis glandularis and 23 cases of normal bladder mucosa, and the expression of AKO62675 was compared with that in 40 cases of bladder cancer tissue, 15 cases of cystitis glandularis and 23 cases of normal bladder mucosa. To investigate the expression of this fragment in bladder tumor tissue, cystitis glandularis tissue and normal bladder mucosa, and analyze whether the fragment is related to the occurrence and development of bladder cancer. Results: 1, the analysis found that the sequence is different from the specific position of CK20 on the chromosome. NCBI was submitted to obtain the gene number: KR780251 and the protein number: AKO62675.2. The polyclonal antibody: Anti-KR was detected by ELISA method. The titer of the polyclonal antibody was 1: 64000, which met the detection requirements. Immunohistochemical staining of cystitis glandularis and normal bladder tissue: the positive rate of Anti-KR staining was 34.78 in normal bladder mucosa, 100.00in bladder cancer tissue and Anti-KR staining in cystitis glandularis tissue. It was 100.00g, but the staining was lighter than that of bladder cancer. Conclusion the full length KR780251 of the 1: 1 EST fragment has 1839bp. the protein AKO62675 encoded by ORFN KR780251 may be a mutant of normal bladder tissue gene .2AKO62675, which is highly expressed in bladder cancer.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.14

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本文編號：1849074