摘要
目的:筛选与人小细胞肺癌转移相关基因,揭示肺癌的转移机制。方法:采用激光俘获显微切割(LCM)及mRNA差异显示技术(DD)对手术切除的小细胞肺癌、癌转移淋巴结进行筛选并克隆与转移相关的基因片段,测定其序列后在基因库(GenBank)中进行BLAST检索分析其同源性。结果:小细胞肺癌原发灶与癌转移淋巴结的基因表达具有明显差异,共获得差异片段(EST,表达序列标签)24个,选择差异明显的3个片段克隆测序后,发现3片段均位于肿瘤高度相关的3q、7q和10q,均为未知序列,未发现同源基因,可能为新的基因片段。用RT-PCR对位于抑癌基因PTEN上游的L1片段进行验证的结果证实了该片段在小细胞肺癌的高表达率,具有统计学意义(P<0.05)。结论:人小细胞肺癌原发灶与癌转移淋巴结的基因表达存在差异,所测序的3个基因片段均为未知基因序列,这些片段可能为小细胞肺癌的转移相关候选基因,其序列和功能值得进一步深入研究。应用DDRT-PCR技术有望找到调控肿瘤转移的基因,为肿瘤的诊断和治疗提供新的线索。
Objective： To find the metastasis-related genes in small cell lung cancer （SCLC）, and to investigate the mechanism of SCLC metastasis. Methods： Laser capture microdissection （LCM） and fluoro-differential display reverse transcriptase polymerase chain reaction （fluoro-DDRT-PCR） were performed to screen differentially expressed gene fragments between primary SCLC and metastatic lymph nodes. The sequences of gene fraganents were subsequently analyzed and compared to known sequences in Genbank. Resuits： Twenty-four cDNA fragments （expressed sequence tags, ESTs） which differentially expressed in primary SCLC and metastatic lymph nodes were isolated by using fluoro-DDRT-PCR. Three of these ESTs were cloned, and the sequencing analysis of these clones showed that they were novel gene fragments which had no homology with known genes in GenBank. The expression rate of L1 gene fragment which located near tumor suppressor gene PTEN in primary SCLC was significant higher than that in metastatic lymph nodes （ P 〈 0.05）. Conclusion： The gene expression is different between primary SCLC and the metastatic lymph nodes. These 3 novel gene fragments may be candidates of SCLC metastasis-related genes. The results of this study may provide novel points of therapeutic intervention for SCLC.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2006年第1期31-33,共3页
Journal of China Medical University
基金
国家自然科学基金资助项目(30170194)
关键词
激光俘获显微切割
MRNA差异显示
转移相关基因
小细胞肺癌
laser capture microdissection
mRNA differential display
metastasis-related genes
small cell lung cancer
