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[目的]通过激光捕获显微切割技术(LCM)构建喉部纯化组织差异基因表达谱,筛选与进展期喉癌相关的候选靶基因。[方法]采用激光捕获显微切割技术纯化分离8例声门型喉癌组织标本中的癌细胞及对应癌旁组织的黏膜上皮细胞,结合HGU133.Plus2.0芯片技术,构建16例喉部纯化组织标本差异基因组表达谱,筛选出进展期喉癌相关靶基因;采用QRT-PCR检测特异性的候选靶基因的表达情况。[结果]将不同分期的喉癌组与癌旁正常上皮组基因表达谱分别进行组内、组间比较,共筛选出与喉癌临床进展相关的候选靶基因761个;对部分特异性基因如KRT19、KRT23基因进行mRNA水平的检测,其结果与基因表达谱的结果具有一致性。[结论]利用激光捕获显微切割结合寡核苷酸芯片技术构建喉癌差异基因表达谱,可准确、高效地筛选出与喉癌临床进展相关的候选靶基因。
[Objective] To construct the differentially expressed gene expression profile of laryngeal tissues by laser capture microdissection (LCM) and screen the candidate target genes related to advanced laryngeal carcinoma. [Method] The laser capture microdissection technique was used to purify the cancer cells in the glottic laryngeal carcinoma of 8 cases and the corresponding mucosal epithelial cells in paracancerous tissues. Combined with HGU133.Plus2.0 chip technology, 16 cases of laryngeal purification Tissue samples were differentially expressed in genomic DNA and the target genes of advanced laryngeal carcinoma were screened out. The expression of specific target genes was detected by QRT-PCR. [Results] The gene expression profiles of laryngeal cancer group and adjacent non-cancerous normal epithelium group were compared respectively in groups and between groups. 761 candidate target genes related to the clinical progress of laryngeal cancer were screened out. The expression of some specific genes KRT19, KRT23 gene mRNA level detection, the results and gene expression profiles of the results are consistent. [Conclusion] By using laser capture microdissection and oligonucleotide microarray technology to construct differential expression profiles of laryngeal carcinoma, we can screen out the target genes that are relevant to the clinical progress of laryngeal carcinoma accurately and efficiently.