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背景:宫颈癌是女性最常见的恶性肿瘤之一,其发生、发展与环加氧酶2密切相关。目的:探讨人宫颈癌组织中是否存在环加氧酶2选择性剪接异构体的表达及其可能意义。设计:非随机对照实验。单位:重庆医科大学生物化学与分子药理学重点实验室,重庆医科大学附属第一医院妇产科。对象:选择13例重庆医科大学附属第一医院妇产科2002-03/2002-04宫颈癌住院患者的癌组织及正常组织。方法:设计一对全新引物,并采用反转录聚合酶链反应技术,从宫颈癌组织中分离环加氧酶2mRNA,然后对电泳条带进行克隆、测序和分析。主要观察指标:①观察癌组织与正常组织反转录聚合酶链反应琼脂糖电泳结果。②分析癌组织与正常组织电泳条带测序结果。结果:①正常宫颈组织未发现环加氧酶2目的条带(252bp),而宫颈癌组织则出现两条条带,除环加氧酶2目的条带外,还有一新的电泳带(534bp),经克隆和测序证实,该新条带除目的条带的环加氧酶2DNA的第7、8外显子序列外,还包含第7、8外显子间的内含子序列,即保留的内含子(282bp)。②根据阅读框推测,在保留的内含子第48～50碱基出现终止密码。结论:发现宫颈癌组织中存在环加氧酶2基因的选择性剪接异构体(Genbankaccessionnumber:BU493602),其所编码蛋白可能较已报道的环加氧酶2酶蛋白小。
Background: Cervical cancer is one of the most common malignant tumors in women. Its occurrence and development are closely related to cyclooxygenase 2. Objective: To investigate the expression of cyclooxygenase 2 alternative splicing isoform in human cervical carcinoma and its possible significance. Design: Non-randomized controlled experiment. Unit: Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing Medical University, Obstetrics and Gynecology, First Affiliated Hospital of Chongqing Medical University. PARTICIPANTS: 13 cases selected from the First Affiliated Hospital of Chongqing Medical University Obstetrics and Gynecology 2002-03 / 2002-04 Cancer patients and normal tissues of inpatients with cervical cancer. Methods: A pair of new primers were designed and cyclooxygenase 2 mRNA was isolated from cervical cancer tissues by reverse transcription polymerase chain reaction (PCR). Then the electrophoresis bands were cloned, sequenced and analyzed. MAIN OUTCOME MEASURES: ①To observe the reverse transcriptase polymerase chain reaction agarose gel electrophoresis results of cancer tissues and normal tissues. ② analysis of cancer tissue and normal tissue electrophoresis band sequencing results. Results: (1) No band of 252bp was detected in normal cervical tissues, while two bands appeared in cervical cancer tissues. In addition to the 2 band of cyclooxygenase, a new band of 534bp ), Confirmed by cloning and sequencing, this new band in addition to the purpose of the band of cyclooxygenase 2DNA 7,8 exon sequence, also contains the 7th, exon 8 intron sequence, that Reserved intron (282 bp). ② According to the reading frame speculation, in the reserved intron 48 ~ 50 base stop codon. CONCLUSION: The genomic accession number (BU493602) of cyclooxygenase 2 gene is found in cervical cancer tissues, which may be smaller than the reported cyclooxygenase 2 enzyme protein.