目的探讨宫颈癌染色体3p14区域D3S1300、D3S1600位点微卫星不稳定性(microsatellite instability,MSl)及等位基因杂合性缺失(loss of heterozygosity,LOH)频率,为准确定位宫颈癌相关肿瘤抑制基因位点提供实验依据,并探讨LOH及MSI与宫颈...目的探讨宫颈癌染色体3p14区域D3S1300、D3S1600位点微卫星不稳定性(microsatellite instability,MSl)及等位基因杂合性缺失(loss of heterozygosity,LOH)频率,为准确定位宫颈癌相关肿瘤抑制基因位点提供实验依据,并探讨LOH及MSI与宫颈癌临床分期、病理分级的相关性。方法选择3p14区域两个微卫星多态性位点,显微分离提取41例宫颈癌石蜡切片中的正常组织和肿瘤组织,经PCR扩增及聚丙烯酰胺凝胶电泳和硝酸银染色,进行LOH及MSI研究。结果41例样本中有25例至少存在一个位点的LOH,D3S1300位点LOH的频率为35%,D3S1600位点LOH的频率为28%。MSI发生频率相对较低,D3S1300的MSI频率为7.5%,D3S1600的MSI频率为12.8%。D3S1300、D3S1600位点LOH的发生率与临床分期、病理分级相关性有显著意义(P<0.05),而MSI与之无显著的差别(P>0.05)。结论3p14区域内D3S1300、D3S1600位点具有较高的LOH,提示这两个微卫星位点附近可能存在尚未克隆的与宫颈癌发生、发展相关的肿瘤抑制基因。宫颈癌染色体3p14区域LOH与临床分期、病理分级成正相关,提示检测该区域的LOH可作为病程进展及预后的重要参考指标。MSI在本研究中与宫颈癌临床分期、病理分级无明显相关。展开更多
Objective To identify novel tumor suppressor genes at chromosome 3p24-26 in human nasopharyngeal carcinoma (NPC).Methods Twenty epithelial-derived expressed sequence tags (EST) were selected from chromosome 3p24-26. R...Objective To identify novel tumor suppressor genes at chromosome 3p24-26 in human nasopharyngeal carcinoma (NPC).Methods Twenty epithelial-derived expressed sequence tags (EST) were selected from chromosome 3p24-26. RT-PCR and Northern blot were used to detect the expression of the ESTs in NPC cell line, HNE-1, and primary cultures of normal nasopharyngeal epithelial cells. One EST, which was substantially downregulated in the HNE-1 cell line, was detected in 19 NPC biopsy samples, cDNA library screening was used to get its full sequence and the sequence of this novel gene was analyzed.Results A novel gene located at chromosome 3p25.3 was obtained and named NAG-7. It was downregulated in 26.3％ (5/19) of NPC biopsy samples. Its 1677 bp full length cDNA had a potential open reading frame predicting a 94 amino acid protein with a molecular weight of 11023.87 Dalton. Analysis of the NAG-7 gene showed that it was a transmembrane protein containing a protein kinase C phosphorylation site and a myristyl site. It has no significant homology to any reported genes in the database of GenBank. Conclusion NAG-7 is a novel gene downregulated in NPC, suggesting that it may be involved in the development of NPC.展开更多
文摘目的探讨宫颈癌染色体3p14区域D3S1300、D3S1600位点微卫星不稳定性(microsatellite instability,MSl)及等位基因杂合性缺失(loss of heterozygosity,LOH)频率,为准确定位宫颈癌相关肿瘤抑制基因位点提供实验依据,并探讨LOH及MSI与宫颈癌临床分期、病理分级的相关性。方法选择3p14区域两个微卫星多态性位点,显微分离提取41例宫颈癌石蜡切片中的正常组织和肿瘤组织,经PCR扩增及聚丙烯酰胺凝胶电泳和硝酸银染色,进行LOH及MSI研究。结果41例样本中有25例至少存在一个位点的LOH,D3S1300位点LOH的频率为35%,D3S1600位点LOH的频率为28%。MSI发生频率相对较低,D3S1300的MSI频率为7.5%,D3S1600的MSI频率为12.8%。D3S1300、D3S1600位点LOH的发生率与临床分期、病理分级相关性有显著意义(P<0.05),而MSI与之无显著的差别(P>0.05)。结论3p14区域内D3S1300、D3S1600位点具有较高的LOH,提示这两个微卫星位点附近可能存在尚未克隆的与宫颈癌发生、发展相关的肿瘤抑制基因。宫颈癌染色体3p14区域LOH与临床分期、病理分级成正相关,提示检测该区域的LOH可作为病程进展及预后的重要参考指标。MSI在本研究中与宫颈癌临床分期、病理分级无明显相关。
文摘Objective To identify novel tumor suppressor genes at chromosome 3p24-26 in human nasopharyngeal carcinoma (NPC).Methods Twenty epithelial-derived expressed sequence tags (EST) were selected from chromosome 3p24-26. RT-PCR and Northern blot were used to detect the expression of the ESTs in NPC cell line, HNE-1, and primary cultures of normal nasopharyngeal epithelial cells. One EST, which was substantially downregulated in the HNE-1 cell line, was detected in 19 NPC biopsy samples, cDNA library screening was used to get its full sequence and the sequence of this novel gene was analyzed.Results A novel gene located at chromosome 3p25.3 was obtained and named NAG-7. It was downregulated in 26.3％ (5/19) of NPC biopsy samples. Its 1677 bp full length cDNA had a potential open reading frame predicting a 94 amino acid protein with a molecular weight of 11023.87 Dalton. Analysis of the NAG-7 gene showed that it was a transmembrane protein containing a protein kinase C phosphorylation site and a myristyl site. It has no significant homology to any reported genes in the database of GenBank. Conclusion NAG-7 is a novel gene downregulated in NPC, suggesting that it may be involved in the development of NPC.
