Tan Xiaojie, Zhai Yujia, Chang Wenjun, Hou Jianguo, He Songqin, Lin Liping, Yu Yongwei, Xu Danfeng, Xiao Jianru, Ma Liye, Wang Guoping, Cao Tinghu, Cao Guangwen
Department of Epidemiology, Second Military Medical University, Shanghai, China.
Int J Cancer. 2008 Sep 1;123(5):1080-8. doi: 10.1002/ijc.23637.
Metastatic clear-cell renal-cell carcinoma (ccRCC) has a poor prognosis and unpredictable course, and there are no molecular markers that reliably predict ccRCC metastasis. In this study, ccRCC specimens from 84 patients were directly cultured in vitro. Primary cultures from 38 of 94 specimens contained more than 90% tumor cells at the fourth passage. After identification by immunostaining, the primary cultures of metastatic and nonmetastatic ccRCC specimens from the age- and gender-matched patients were subjected to cDNA microarray assays. A total of 842 differentially expressed genes with a FDR (false discovery rate) of 4.79% were identified. Pathway analysis and co-occurrence with "cancer", "metastasis" and "invasion" in the literature annotations functionally enriched the 842 genes and provided an indication of the reliability of our microarray assays. Novel genes associated with metastasis were selected based on protein-protein interactions between 205 differentially expressed genes that co-occurred with "metastasis" and those that did not co-occur with "metastasis" on Medline, and the results of co-expression analysis between the co-occurred genes and unpublished genes. FSTL1, AV722783, SLC15A1, DDX17, ORC2L and PKMYT1 were found to be potential ccRCC metastasis-associated novel genes, according to expression patterns in cultures and tumor tissues. Interestingly, the upregulated genes (CAV1, PKMYT1 and ORC2L) were also upregulated and the downregulated genes (FSTL1, GSTM3, CYR61, SLC15A1 and AV722783) were also downregulated in the primary ccRCC specimens compared with expression in adjacent renal tissues in 37 patients. This study has identified new candidate biomarkers and targets for the early diagnosis and treatment of ccRCC metastasis.
转移性透明细胞肾细胞癌（ccRCC）预后较差且病程不可预测，目前尚无可靠预测ccRCC转移的分子标志物。在本研究中，对84例患者的ccRCC标本进行体外直接培养。94个标本中的38个原代培养物在第4代时肿瘤细胞含量超过90%。经免疫染色鉴定后，对年龄和性别匹配患者的转移性和非转移性ccRCC标本的原代培养物进行cDNA微阵列分析。共鉴定出842个差异表达基因，错误发现率（FDR）为4.79%。通路分析以及与文献注释中“癌症”“转移”和“侵袭”的共现情况对这84个基因进行了功能富集，并表明了我们微阵列分析的可靠性。基于205个与“转移”共现的差异表达基因和未与“转移”共现的差异表达基因之间的蛋白质-蛋白质相互作用，以及共现基因与未发表基因之间的共表达分析结果，筛选出与转移相关的新基因。根据培养物和肿瘤组织中的表达模式，发现FSTL1、AV722783、SLC15A1、DDX17、ORC2L和PKMYT1是潜在的与ccRCC转移相关的新基因。有趣的是，与37例患者相邻肾组织中的表达相比，原发性ccRCC标本中上调的基因（CAV1、PKMYT1和ORC2L）也上调，而下调的基因（FSTL1、GSTM3、CYR61、SLC15A1和AV722783）也下调。本研究确定了用于ccRCC转移早期诊断和治疗的新候选生物标志物和靶点。
