Macrodissection versus microdissection of rectal carcinoma: minor influence of stroma cells to tumor cell gene expression profiles.
Macrodissection versus microdissection of rectal carcinoma: minor influence of stroma cells to tumor cell gene expression profiles.
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DOI：
10.1186/1471-2164-6-142
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发表时间：
2005-10-14
期刊：
影响因子：
4.4
通讯作者：
Peltenburg LT
中科院分区：
文献类型：
--
作者：
de Bruin EC;van de Pas S;Lips EH;van Eijk R;van der Zee MM;Lombaerts M;van Wezel T;Marijnen CA;van Krieken JH;Medema JP;van de Velde CJ;Eilers PH;Peltenburg LT
通过微阵列分析同时比较数千个基因，可以推断出致癌、肿瘤进展和患者预后的分子决定因素。然而，手术切除的癌组织中存在的基质细胞可能会掩盖这些样本的肿瘤细胞特异性基因表达谱。为了避免这种并发症，可以进行激光显微切割以将肿瘤上皮与周围的基质和健康组织分离。在这份报告中，我们比较了RNA分离的macrosuttered，其中只有周围的健康组织已被删除，和microdissected直肠癌样本的微阵列分析，以确定最可靠的方法来检测肿瘤细胞衍生的基因表达的微阵列分析。由于显微切割产生的组织和RNA量较低，因此需要额外的mRNA扩增轮次以获得足够的RNA用于微阵列实验。这些第二轮扩增影响基因表达谱。此外，基质细胞在宏观解剖样品中的存在对肿瘤细胞基因表达谱的贡献较小，这可以通过观察到从肿瘤上皮细胞提取的RNA多于从基质提取的RNA来解释。这些数据表明，更方便的宏观解剖程序可以充分使用，并产生可靠的数据，关于肿瘤细胞特异性基因表达谱的鉴定。
The molecular determinants of carcinogenesis, tumor progression and patient prognosis can be deduced from simultaneous comparison of thousands of genes by microarray analysis. However, the presence of stroma cells in surgically excised carcinoma tissues might obscure the tumor cell-specific gene expression profiles of these samples. To circumvent this complication, laser microdissection can be performed to separate tumor epithelium from the surrounding stroma and healthy tissue. In this report, we compared RNAs isolated from macrodissected, of which only surrounding healthy tissue had been removed, and microdissected rectal carcinoma samples by microarray analysis in order to determine the most reliable approach to detect the expression of tumor cell-derived genes by microarray analysis. As microdissection yielded low tissue and RNA quantities, extra rounds of mRNA amplification were necessary to obtain sufficient RNA for microarray experiments. These second rounds of amplification influenced the gene expression profiles. Moreover, the presence of stroma cells in macrodissected samples had a minor contribution to the tumor cell gene expression profiles, which can be explained by the observation that more RNA is extracted from tumor epithelial cells than from stroma. These data demonstrate that the more convenient procedure of macrodissection can be adequately used and yields reliable data regarding the identification of tumor cell-specific gene expression profiles.
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影响因子：
14.9
作者：
Baugh, L. R.;Hill, A. A.;Hunter, Craig P.
通讯作者：
Hunter, Craig P.
影响因子：
4.4
作者：
Hu, LM;Wang, J;Zhang, W
通讯作者：
Zhang, W
影响因子：
2.7
作者：
Feldman, AL;Costouros, NG;Libutti, SK
通讯作者：
Libutti, SK
DOI：
10.1073/pnas.96.16.9212
发表时间：
1999-08-03
影响因子：
11.1
作者：
Perou, CM;Jeffrey, SS;Botstein, D
通讯作者：
Botstein, D
DOI：
10.1073/pnas.2536479100
发表时间：
2004-01-13
影响因子：
11.1
作者：
Stuart, RO;Wachsman, W;Mercola, D
通讯作者：
Mercola, D