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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。
Objective To investigate the frequency of microsatellite instability (MSl) and allele loss of heterozygosity (LOH) at D3S1300 and D3S1600 loci in chromosome 3p14 of cervical cancer and to determine the location of tumor suppressor genes related to cervical cancer Point to provide experimental evidence, and to explore the correlation between LOH and MSI and clinical stage and pathological grade of cervical cancer. Methods Two microsatellite loci in 3p14 region were selected. The normal tissues and tumor tissues in 41 cases of cervical cancer paraffin sections were extracted by micro-isolation. The PCR products were amplified by PCR and polyacrylamide gel electrophoresis and silver nitrate staining. And MSI research. Results Of the 41 samples, there were at least one LOH in 25 cases, 35% of LOH in D3S1300 and 28% of LOH in D3S1600. The frequency of MSI is relatively low with an MSI frequency of 7.5% for the D3S1300 and 12.8% for the D3S1600. The incidence of LOH at D3S1300 and D3S1600 sites was significantly correlated with clinical stage and pathological grade (P <0.05), while there was no significant difference between MSI and SLS (P> 0.05). Conclusion There is a high LOH in D3S1300 and D3S1600 sites in 3p14 region, suggesting that there may exist some tumor suppressor genes that have not been cloned and related to the occurrence and development of cervical cancer near the two microsatellite loci. Cervical cancer chromosome 3p14 region LOH and clinical stage, pathological grade is positively correlated, suggesting that the detection of LOH in the region can be used as an important reference course of disease progression and prognosis. MSI in this study and cervical cancer clinical stage, pathological grade no significant correlation.