Exploration of biomarkers for lymph node metastasis in patients with endometrial cancer using exon-expression microarray.
Exploration of biomarkers for lymph node metastasis in patients with endometrial cancer using exon-expression microarray.
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DOI：
10.1200/jco.2011.29.15_suppl.5100
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发表时间：
2011-05
期刊：
影响因子：
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通讯作者：
S. Sudo;Y. Konno;T. Odagiri;T. Kato;M. Hosaka;M. Takeda;H. Watari;M. Kaneuchi;N. Sakuragi
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文献类型：
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作者：
S. Sudo;Y. Konno;T. Odagiri;T. Kato;M. Hosaka;M. Takeda;H. Watari;M. Kaneuchi;N. Sakuragi
5100背景：为了在术前利用原发癌组织预测淋巴结转移的可能性，我们探索了子宫内膜癌患者淋巴结转移的假定生物标志物基因。方法从2001年8月至2010年12月在我院行子宫切除术、双侧输卵管卵巢切除术、盆腔及腹主动脉旁淋巴结清扫术患者的原发癌组织中提取mRNA。Affytron Exon Microarray分选出在淋巴结阴性和阳性组之间显示显著差异表达的转录本。为了验证微阵列结果，采用实时PCR和Taqman（R）基因表达测定。27例和61例乳腺样腺癌的mRNA样本分别用于微阵列和实时PCR。对于微阵列结果的统计分析，使用ArrayAssist 5.0。结果ANKRD 36（ankyrin repeat domain containing 36）、VPS 13 A、ZNF 577、CROP（cisplatin resistance-associated overexpressed protein）、MALAT-1（metastasis-associated lung adenocarcinoma transcript 1）和TIMP 3（tissue inhibitor of metalloproteinase 3）6种转录本的表达水平在淋巴结阴性组和淋巴结阳性组间差异有统计学意义。为了验证，实时PCR将6个候选物缩小到4个在淋巴结阳性样品中上调的转录物; ANKRD 36、VPS 13 A、CROP和MALAT-1。使用定制的针对ANKRD 36蛋白的肽抗体和市售的抗CROP抗体，进行免疫染色分析。有趣的是，石蜡包埋的癌组织中ANKRD 36和CROP的免疫染色强度对应于实时PCR分析的mRNA表达水平。转移淋巴结ANKRD 36和CROP均呈阳性表达。结论：我们在淋巴结阳性的子宫内膜癌原发灶中发现了4种上调的转录本。它们可能是子宫内膜癌中假定的淋巴结阳性生物标志物。
5100 Background: To predict possibility of lymph node metastasis using primary cancer tissue preoperatively, we explored putative biomarker genes of node metastasis in patients with endometrial cancer. METHODS mRNAs were extracted from primary cancer tissues of patients underwent hysterectomy, bilateral salpingo-oophorectomy with pelvic and para-aortic lymphadenectomy in our institute from August 2001 to December 2010. Affymetrix Exon Microarray sorted out transcripts showing significantly different expression between node-negative and node-positive groups. To validate microarray results, realtime PCR with Taqman(R) Gene Expression Assays were employed. Twenty-seven and 61 mRNA samples with endometrioid adenocarcinoma were used for microarray and realtime PCR, respectively. For statistical analyses of microarray results, ArrayAssist 5.0 was used. RESULTS Expression levels of 6 transcripts, ANKRD36 (ankyrin repeat domain containing 36), VPS13A, ZNF577, CROP (cisplatin resistance-associated overexpressed protein), MALAT-1 (metastasis-associated lung adenocarcinoma transcript 1) and TIMP3 (tissue inhibitor of metalloproteinase 3) showed significant differences between node-negative and node-positive groups in microarray analyses. For the validation, realtime PCR narrowed down 6 candidates to 4 upregulated transcripts in node-positive samples; ANKRD36, VPS13A, CROP and MALAT-1. Using custom-made peptide antibody against ANKRD36 protein and commercially available anti-CROP antibody, immunostaining analyses were performed. Interestingly, immunostaining intensity for ANKRD36 and CROP in paraffin-embedded carcinoma tissues corresponded to mRNA expression levels of realtime PCR analyses. Metastatic lymph node also showed positive for both of ANKRD36 and CROP. CONCLUSIONS We have identified 4 upregulated transcripts in primary cancer focus of node-positive endometrial cancer. They might be the putative node-positive biomarkers in endometrial cancer.