Sen Sudip, Ateeq Bushra, Sharma Himani, Datta Palika, Gupta Siddhartha Datta, Bal Sabyasachi, Kumar Arvind, Singh Neeta
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, 110029, India.
Life Sci. 2008 Mar 26;82(13-14):772-9. doi: 10.1016/j.lfs.2008.01.009. Epub 2008 Jan 30.
Early metastasis and a poor five-year survival make lung cancer the leading cause of cancer related deaths worldwide. The clinical profile of lung cancer patients in India differs from the West as they present earlier, with squamous cell carcinoma being the commonest histological type. We compared gene expression profiles in primary lung squamous cell carcinoma (LSCC) and matched normal lung tissues in Asian Indians. Using suppression subtractive hybridization, two subtracted cDNA libraries containing differentially expressed genes in the tumors were constructed. Differential expression was confirmed by reverse Northern blot analysis. DNA of confirmed clones was sequenced and subjected to GenBank Blast searches. RNA expression levels were then analyzed by Northern blotting and validated by semiquantitative RT-PCR (in 10 cases of NSCLC). Seventeen differentially expressed gene cDNA fragments of LSCC were analyzed. The differentially expressed genes included those associated with cellular metabolism, cell-cycle, -structure, -adhesion, transcription, proliferation, apoptosis and signal transduction. The study provided first evidence that KIAA0767, a Death Inducing Protein, a novel p53 independent target of E2F1, and Geminin, an inhibitor of DNA replication are differentially expressed in LSCC. Identification of the differentially expressed genes in lung cancer in this study may serve as better molecular markers for early diagnosis and identifying novel intervention sites for anticancer therapy.
早期转移和较差的五年生存率使肺癌成为全球癌症相关死亡的主要原因。印度肺癌患者的临床特征与西方不同,因为他们就诊更早,鳞状细胞癌是最常见的组织学类型。我们比较了亚洲印度人原发性肺鳞状细胞癌(LSCC)和配对的正常肺组织中的基因表达谱。利用抑制性消减杂交技术,构建了两个包含肿瘤中差异表达基因的消减cDNA文库。通过反向Northern印迹分析确认差异表达。对确认克隆的DNA进行测序并进行GenBank Blast搜索。然后通过Northern印迹分析RNA表达水平,并通过半定量RT-PCR(在10例非小细胞肺癌中)进行验证。分析了17个LSCC差异表达基因的cDNA片段。差异表达的基因包括与细胞代谢、细胞周期、细胞结构、细胞黏附、转录、增殖、凋亡和信号转导相关的基因。该研究首次提供证据表明,一种死亡诱导蛋白KIAA0767(E2F1的一种新型p53非依赖性靶点)和一种DNA复制抑制剂Geminin在LSCC中差异表达。本研究中肺癌差异表达基因的鉴定可能作为更好的分子标志物用于早期诊断,并为抗癌治疗确定新的干预位点。