人spindlin1基因的功能研究

发布时间：2018-01-26 07:55

本文关键词： spindlin1 亚细胞定位成瘤性信号传导　出处：《中国人民解放军军事医学科学院》2005年博士论文　论文类型：学位论文

【摘要】：卵巢癌是目前严重威胁妇女健康的恶性肿瘤,由于缺少早期临床诊断症状,60-70%的卵巢癌处于晚期诊断,手术难以根治,而目前治疗卵巢癌的药物特异性和敏感性均较差,使得卵巢癌在妇科肿瘤中死亡率最高,5年生存率一直徘徊在40%左右。因此寻找在卵巢癌的发生中敏感和特异的基因改变将有可能为临床卵巢癌的诊治提供有意义的生物学指标,为抗癌药物的研究提供新靶点,具有重要的理论和实际意义。我们研究小组采用改良的mRNA差异显示PCR技术对卵巢癌和正常卵巢组织之间的差异表达基因进行了一系列的筛选和鉴定,获得并克隆了一个新基因,由于它与鼠类spindlin基因高度同源,因此命名为人spindlin1基因(GenBank登录号为AF317228)。对其分析如下: 1 新基因cDNA长度为4375bp,基因在236-949bp含有完整开放读框,预测编码由237个氨基酸组成的蛋白质。在新基因cDNA开放读框的第一个ATG上游-126bp处有同框终止密码子TGA,在3’末端4350bp处为典型的加尾信号AATAAA,此后18bp处为polyA尾。新基因cDNA全长序列与鼠的spindlin基因核酸序列具有96%同源性。基因在236-949bp含有完整开放读框,编码237个氨基酸,所编码氨基酸与鼠spindlin基因所编码的241个氨基酸具有98%的同源性。且新基因与鼠spindlin1基因一样、均在3’端具有长的非编码调控序列。即新基因cDNA在核酸序列、编码氨基酸序列和基因结构三方面与鼠的spindlin基因高度同源。Spindin1基因内含子和外显子剪切位点分析表明,所得基因序列与人第9号染色体RP11-317b17克隆序列中的5个部分顺序一致,可以推断基因来源于人第9号染色体。cDNA由5个外显子组成,基因组DNA由五个外显子和4个内含子组成,基因组全长为52.3kb。进一步分析基因组中内含子和外显子的结构特征,表明内含子和外显子之间的剪切完全符合GT-AT法则。 2 spindlin1基因编码蛋白为一亲水性膜内蛋白,分子量约为27Kd,等电点为5.615。它含有三个重复的spin-ssty结构域,属于spin/ssty家族。生物信息学预测其具有MAPK磷酸化位点、酪氨酸蛋白激酶磷酸化位点、RNA结合蛋白作用位点、cAMP依赖性酪氨酸蛋白激酶磷酸化位点、PKC磷酸化位点等。 3 spindlin 1基因的染色体定位分析表明其定位在第9号染色体的长臂9q22.1-22.3
[Abstract]:Ovarian cancer is a serious threat to women's health at present. Due to the lack of early clinical diagnosis symptoms 60% to 70% of ovarian cancer is in the late diagnosis, surgery is difficult to cure. At present, the drug specificity and sensitivity of the treatment of ovarian cancer are poor, making ovarian cancer in gynecological tumors the highest mortality. The 5-year survival rate has been hovering around 40%. Therefore, searching for sensitive and specific gene changes in the pathogenesis of ovarian cancer may provide valuable biological indicators for the diagnosis and treatment of ovarian cancer. It is of great theoretical and practical significance to provide new targets for the study of anticancer drugs. Our team used modified mRNA differential display PCR to screen and identify differentially expressed genes between ovarian cancer and normal ovarian tissues. A novel gene was obtained and cloned because of its high homology with mouse spindlin gene. Therefore, the accession number of human spindlin1 gene GenBank was AF3172281.The analysis was as follows: 1 the cDNA length of the new gene was 4375bp.The gene contained a complete open reading frame in 236-949bp. The predicted protein is composed of 237 amino acids. There is a codon TGA in the first ATG upstream -126 BP of the open reading frame of the new gene cDNA. A typical caudal signal AATAAA was found at 4350bp at the 3'terminal. The whole cDNA sequence of the new gene has 96% homology with the nucleic acid sequence of mouse spindlin gene, and the gene contains complete opening in 236-949 BP. Read the box. It encodes 237 amino acids and has 98% homology with 241 amino acids encoded by mouse spindlin gene. The new gene is the same as mouse spindlin1 gene. Both of them have long non-coding regulatory sequences at the 3 'end, that is, the new gene cDNA is in nucleic acid sequence. The amino acid sequence and gene structure were highly homologous to mouse spindlin gene. The analysis of intron and exon shear site of Spindin1 gene showed that the amino acid sequence and gene structure were highly homologous to the mouse spindlin gene. The obtained gene sequence is consistent with the five partial sequences of human chromosome 9 RP11-317b17 clone sequence. It can be concluded that the gene originated from human chromosome 9. The gene consists of five exons. Genomic DNA consists of five exons and four introns, and the length of the genome is 52.3 kb. the structural characteristics of introns and exons in the genome are further analyzed. It is shown that the shear between intron and exon conforms to the GT-AT rule. 2 spindlin1 gene is a hydrophilic intramembrane protein, and its molecular weight is about 27kd. The isoelectric point is 5.615. It contains three repeated spin-ssty domains, belonging to the spin/ssty family. Bioinformatics predicts that it has MAPK phosphorylation sites. The phosphorylation site of tyrosine protein kinase and the phosphorylation site of RNA-binding protein are camp dependent tyrosine protein kinase phosphorylation site and PKC phosphorylation site. Chromosome mapping analysis of 3 spindlin 1 gene showed that it was located in the long arm 9q22.1-22.3 of chromosome 9.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R341

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