Li Yalun, He Yanqi, Qiu Zhixin, Zhou Bin, Shi Shaoqin, Zhang Kui, Luo Yangkun, Huang Qian, Li Weimin
Department of Respiratory Medicine, West China Hospital of Sichuan University, 37 Guoxue Xiang, Chengdu, Sichuan, 610041, China.
Tumour Biol. 2014 Dec;35(12):11719-26. doi: 10.1007/s13277-014-2011-0. Epub 2014 Sep 26.
Accumulating evidence supports that genetic factors are another risk factors for lung cancer. Previously, we used whole exome sequencing with sanger sequencing to search for genetic-related mutations in one of four individuals from a pedigree with lung cancer history. Then, we used PCR-RFLP and direct-sequence in the sample size of 318 individuals with lung cancer (cases) and 272 controls. Recently, we detected two new genes including CRTC2 (CREB regulated transcription coactivator 2) and PROM1(human prominin-1,CD133). We investigated the CRTC2 mutation and PROM1 mutation of surgically resected NSCLC tissues (n=200). The presence or absence of CRTC2 and PROM1 mutation was analyzed by direct sequencing. The expression of CRTC2 and PROM1 was studied by western blot and immunohistochemical analysis of the lung cancer tissues which had the mutation of the two genes(cases), the samples without mutations(controls) and the normal lung tissue(controls). CRTC2 and PROM1 mutations in 5 NSCLC tissues and 3 NSCLC tissues out of the samples were identified. The positive results were closely correlated with clinicopathological features, such as male gender, adenocarcinoma, smoker status, and older age (≥55). We found that the CRTC2 and PROM1 expression were significantly higher in tissues of NSCLS with mutations than that without mutations and the normal lung tissue. The results imply that the high expression of CRTC2 and PROM1 may play an important role in the development and hereditary of NSCLC.
越来越多的证据支持遗传因素是肺癌的另一类危险因素。此前,我们利用全外显子测序结合桑格测序,在一个有肺癌病史的家系中的四名个体之一中寻找与遗传相关的突变。然后,我们在318例肺癌患者(病例)和272例对照中采用聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)和直接测序法。最近,我们检测到两个新基因,包括CRTC2(CREB调节转录共激活因子2)和PROM1(人类prominin-1,CD133)。我们研究了手术切除的非小细胞肺癌组织(n=200)中的CRTC2突变和PROM1突变。通过直接测序分析CRTC2和PROM1突变的有无。通过蛋白质免疫印迹法以及对有这两个基因突变的肺癌组织(病例)、无突变的样本(对照)和正常肺组织(对照)进行免疫组织化学分析,研究CRTC2和PROM1的表达。在样本中的5例非小细胞肺癌组织和3例非小细胞肺癌组织中鉴定出CRTC2和PROM1突变。阳性结果与临床病理特征密切相关,如男性、腺癌、吸烟状态和老年(≥55岁)。我们发现,有突变的非小细胞肺癌组织中CRTC2和PROM1的表达明显高于无突变组织和正常肺组织。结果表明,CRTC2和PROM1的高表达可能在非小细胞肺癌的发生和遗传中起重要作用。