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首页> 外文期刊>Cancer biology & therapy >Differential gene expression profiling in aggressive bladder transitional cell carcinoma compared to the adjacent microscopically normal urothelium by microdissection-SMART cDNA PCR-SSH.
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Differential gene expression profiling in aggressive bladder transitional cell carcinoma compared to the adjacent microscopically normal urothelium by microdissection-SMART cDNA PCR-SSH.

机译：通过显微解剖-SMART cDNA PCR-SSH，与相邻的显微镜正常尿路上皮相比，侵袭性膀胱移行细胞癌中的差异基因表达谱。

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摘要

Identifying novel and known genes that are differentially expressed in aggressive bladder transitional cell carcinoma (BTCC) has important implications in understanding the biology of bladder tumorigenesis and developing new diagnostic and therapeutic agents. In this study we identified the differential gene expression profiles comparing tumor to the adjacent microscopically normal mucosa by manual microdissection on frozen sections. The RNAs extracted from microdissected tissues were amplified by SMART cDNA PCR technology to generate forward subtractive cDNA library by suppressive subtractive hybridization (SSH). We obtained 376 positive clones, one hundred clones of aggressive BTCC subtracted cDNA library were selected at random and inserts were reamplified by PCR. After differential screening by reverse dot blotting, 73 positive clones, that contend inserts putatively upregulated in aggressive BTCC, were further analysed by DNA sequencing, GenBank and EST database searching. Sequencing results showed that 66 clones stand for 23 known genes and 7 clones for three new EST (Genbank number: DN236875, DN236874 and DN236873). In conclusion, microdissection-SMART cDNA PCR-SSH allowed for an efficient way to identify aggressive BTCC-specific differential expressed genes that may potentially be involved in the carcinogenesis and/or progression of aggressive BTCC. These differentially expressed genes may be of potential utility as therapeutic and diagnostic targets for aggressive BTCC.
机译：鉴定在侵袭性膀胱移行细胞癌（BTCC）中差异表达的新的和已知的基因，对理解膀胱肿瘤的发生生物学和开发新的诊断和治疗剂具有重要意义。在这项研究中，我们通过在冷冻切片上进行手动显微解剖，鉴定了将肿瘤与相邻的显微镜正常黏膜相比的差异基因表达谱。通过SMART cDNA PCR技术扩增从显微解剖组织中提取的RNA，通过抑制性消减杂交（SSH）生成正向消减cDNA文库。我们获得了376个阳性克隆，随机选择了100个侵略性BTCC减去cDNA文库的克隆，并通过PCR扩增了插入片段。通过反向斑点印迹进行差异筛选后，通过DNA测序，GenBank和EST数据库搜索进一步分析了73个阳性克隆，这些克隆主张在侵略性BTCC中假定上调的插入片段。测序结果显示66个克隆代表23个已知基因，而7个克隆代表三个新的EST（基因库编号：DN236875，DN236874和DN236873）。总之，显微解剖-SMART cDNA PCR-SSH提供了一种有效的方法来鉴定攻击性BTCC特异性差异表达基因，这些基因可能与攻击性BTCC的癌变和/或进展有关。这些差异表达的基因可能具有作为攻击性BTCC的治疗和诊断靶标的潜在用途。

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