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首页> 外文期刊>World Journal of Gastroenterology >Inactivation of RASSF1A, the tumor suppressor gene at 3p21.3 in extrahepatic cholangiocarcinoma.
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Inactivation of RASSF1A, the tumor suppressor gene at 3p21.3 in extrahepatic cholangiocarcinoma.

机译：肝外胆管癌中3p21.3的抑癌基因RASSF1A失活。

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AIM: To evaluate the genetic and epigenetic inactivation mechanism of the RASSF1A tumor suppressor gene at 3p21.3 in extrahepatic cholangiocarcinoma. METHODS: RT-PCR was used to investigate the transcriptional expressing and re-expression of RASSF1A. RASSF1A mutation was analyzed with SSCP and selective sequencing. PCR was performed to detect the loss of heterozygosity (LOH) at the region of chromosome 3p21.3. Genomic DNA were modificated bisulfite and the frequency of methylation of CpG islands in RASSF1A promoter were evaluated by methylation specific PCR (MS-PCR). RESULTS: In all 48 samples and one cell lines of extrahepatic cholangiocarcinoma, the RASSF1A mutation is rare (6.12%, 3/49), 33 samples (68.75%) and QBC-939 cell lines (chi(2) = 14.270, P = 0.001>0.01) showed RASSF1A express inactivation with LOH at microsatellite loci D3S4604. Among these 33 samples and QBC-939, 28 of 33 (84.85%) tumor samples and 1 cell lines were methylated for majority of 16 CpGs, the average frequency is 73.42%. CONCLUSION: The data we present suggest that RASSF1A which we have been searching for at 3p21.3 may be one of the key tumor suppressor gene and play an important role in the pathogenesis of extrahepatic cholangiocarcinoma, and the promoter methylation and allelic loss are the major mechanism for inactivation of RASSF1A.
机译：目的：评价肝外胆管癌中RASSF1A抑癌基因3p21.3的遗传和表观遗传失活机制。方法：采用RT-PCR技术检测RASSF1A的转录表达和再表达。用SSCP和选择性测序分析RASSF1A突变。进行PCR以检测染色体3p21.3区域的杂合性（LOH）的丧失。基因组DNA被修饰的亚硫酸氢盐，并通过甲基化特异性PCR（MS-PCR）评估了RASSF1A启动子中CpG岛的甲基化频率。结果：在全部48个样本和一个肝外胆管癌细胞系中，RASSF1A突变罕见（6.12％，3/49），33个样本（68.75％）和QBC-939细胞系（chi（2）= 14.270，P = 0.001> 0.01）显示RASSF1A在微卫星基因座D3S4604上表达了LOH失活。在这33个样本和QBC-939中，对33个肿瘤样本中的28个（84.85％）和1个细胞系进行了甲基化，以检测16个CpG的大多数，平均频率为73.42％。结论：目前的数据表明，我们一直在3p21.3处寻找的RASSF1A可能是关键的抑癌基因之一，并且在肝外胆管癌的发病机理中起着重要作用，启动子甲基化和等位基因丢失是主要的RASSF1A灭活的机制。

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