【作者】 李坤雨；

【导师】 韩泽广；

【作者基本信息】 华东理工大学 ， 生物化学与分子生物学， 2012， 硕士

【摘要】 肝细胞癌(HCC)是世界范围内最常见的癌症之一,其具有浸润和转移的遗传倾向性。实验室前期采用外显子捕获测序技术,利用Illumina Solexa测序平台分析鉴定伴发有门静脉癌栓(PVTT)的10位肝细胞癌病人样本的体细胞突变。PVTT是肝细胞癌肝内转移形式。对10例肝癌病人的癌旁组织、原位癌及门静脉癌栓共29个组织样本(其中一例病人癌栓组织样本降解)的Solexa测序结果进行分析发现在10例肝癌病人的癌组织中有122574个潜在的突变位点。通过设定特定的筛选条件进行筛选过滤,我们选取了381个定位于编码外显子区域的非沉默性突变位点,通过PCR-Sanger测序法进一步验证,最终重新验证了165个非沉默性突变,验证率为43%。在经过验证的165个非沉默突变位点中经常出现碱基的转换和颠换。我们采用RNA干扰的方法评价了91个突变基因对于细胞活性的影响。结果显示,TMEM35、CDK14、HOXA1、ABCA1都是潜在的癌基因,赋予了肝癌细胞生长和浸润的能力。进一步研究发现TMEM35基因编码具有四跨膜结构域的膜蛋白,其在肝癌细胞生长中发挥重要作用。更多还原

【Abstract】 Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and exhibits an inherited predisposition to infiltrate and metastasize. Here, we use exomic sequencing parallel sequences on an Illumina Solexa platform to identify the somatic mutations in ten HCC patients with portal vein tumor thrombus (PVTT), the intrahepatic metastasis. Through sequencing 29 samples of ten HCC patients’non-cancerous livers, primary and PVTT (one of PVTTwas degradation) we found 122574 somatic mutations on ten HCC patients with PVTT.After analyzed according to described method, we selected 381 non-silenced mutations which located on exon for futher test by sanger sequencing. Finally we retested 165 non-silenced mutations, validation rate was 43%. Base transitions and base transversions were frequently found among 165 confirmed non-silenced mutations. We assessed the role of 91 mutated genes in cellular survival via RNA interference; the results suggest that TMEM35, CDK14, HOXA1, ABCA1 could be potential cancer genes that confer growth and infiltration capabilities to HCC cells. For futher study we found TMEM35 gene, which encode a tetraspanin, have an important role in cell growth of hepatoma cells.更多还原