【作者】 高洪梅；

【导师】 王修海； 王培林；

【作者基本信息】 青岛大学 ， 遗传学， 2004， 硕士

【摘要】 目的 检测肿瘤转移抑制基因nm23-H1在原发性肝细胞癌(hepatocellular carcinoma，HCC)中的表达及其基因突变情况，探讨nm23-H1在HCC发生发展中的作用及肝癌细胞转移的分子生物学机制。方法 应用半定量逆转录聚合酶链反应(RT-PCR)技术及单链构象多态法(single strand conformation polymorphism，SSCP)，对30例肝癌组织、30例癌旁组织和16例正常肝组织中nm23-H1基因的表达情况及cDNA基因突变情况进行检测和分析。结果 30例肝癌患者的癌组织、相应癌旁组织及正常组织中nm23-H1基因阳性表达率均为100％，nm23-H1基因阳性表达率在这三者之间无差异；②nm23-H1基因在肝癌组织、癌旁组织、正常组织中的表达水平分别为1.619±0.407((?)±s)、1.302±0.304((?)±s)及1.232±0.386((?)±s)。统计学分析表明在这三种组织中nm23-H1基因的表达水平有显著差异(P＜0.01)。肝癌组织中nm23-H1基因的表达水平明显低于癌旁组织(P＜0.05)和正常组织(P＜0.01)。该基因在癌旁组织和正常组织中的表达水平无显著差异(P＞0.05)；③低分化肝癌组织中nm23-H1基因表达水平低于高、中分化肝癌组织中nm23-H1基因表达水平(P＜0.05)；④有卫星转移灶的和无卫星灶的肝癌组织中nm23-H1基因表达水平无显著差异(P＞0.05)；⑤肝癌组织中nm23-H1基因表达水平与患者的年龄、性别无显著相关性(P值均大于0.05)；⑥30例肝癌组织及相应的癌旁组织中未发现1例存在nm23-H1 cDNA基因突变。结论 ①原发性肝细胞癌中nm23-H1基因阴性表达率很低；②nm23-H1基因在肝癌组织中的表达水平明显低于癌旁组织和正常组织，提示nm23-H1失活与肝癌发生相关；③原发性肝细胞癌中nm23-H1表达水平与肝癌恶性程度有明显相关性，提示nm23-H1变异可以作为患者预后的指标；④肝癌组织中nm23-H1表达水平与卫星转移灶的有无及患者的性别、年龄均无明显相关性；⑤肝癌组织及癌旁组织中nm23-H1 cDNA基因突变率很低。更多还原

【Abstract】 Objective To detect the expression of nm23-H1 and its cDNA gene mutation in hepatocellular carcinoma (HCC), and study the role of nm23-H1 in the pathogenesis of HCC and elucidate the molecular biology mechanism of hepatocellular carcinoma metastasis. Methods Using semi-quantitive reverse transcriptase-polymerase chain reaction (RT-PCR) and single strand conformation polymorphism (SSCP) , the expression variation and cDNA gene mutation of nm23-Hl gene were detected in 30 HCC tissues, their corresponding adjacent liver tissues and in normal liver tissues of 16 non-liver disease patients. Results 漏the expression rate of nm23-Hl RNA was 100% in the HCC tissues , their corresponding adjacent tissues and in the normal liver tissues. There were no significant difference in the expression rate of nm23-H1 gene among the three kind of tissues(P>0. 05); (2)The expression level of nm23-Hl gene in the HCC tissues, their adjacent tissues and normal liver tissues was 1.619 0.407( x s), 1.302 0.343( x s) and 1.232 0.386( x s), respectively. The expression level of nm23-H1 gene in the HCC tissues was singnificantly lower than that in the adjacent and normal liver tissues (P <0.01); (3) The expression level of nm23-H1 gene in the low-differentiated HCC was much lower than that in the high and middle differentiated HCC tissues (P< 0.01); (4) The expression level of nm23-Hl gene in HCC tissues with satellite focuses was not significantly different from that without satellite focuses (P > 0. 05) ; (5) The expression quantity of nm23-H1 gene in HCC tissues was not closely related to HCC patients’ age and gender (P>0. 05) ; (6) None of the nm23-Hl gene mutation was found in all the hepatocellular carcinoma samples. Conclusion (1)The negative expression rate of nm23-H1 in the HCC tissues is very low; (2) The expression level of nm23-H1 in the HCC tissues is much lower than that in the adjacent and normal liver tissues, which indicates that the inactivation of nm23-H1 is associated with thepathogenesis of HCC; (3)The expression level of nm23-Hl gene is highly associated with the pathological differentiation of HCC, which indicates that the alternation of nm23-Hl can serve as a prognostic indicators in the progression of HCC; (4)The expression level of nm23-Hl gene in HCC tissues was not closely related to HCC patients’ age, gender and whether there are satellite focuses in HCC tissues; (5) The cDNA gene mutation rate of nm23-Hl in the HCC tissues is very low.更多还原