Identification of molecular markers for metastasis-related genes in primary breast cancer cells
Identification of molecular markers for metastasis-related genes in primary breast cancer cells
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DOI：
10.1007/s10585-005-4417-y
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发表时间：
2005-01-01
影响因子：
4
通讯作者：
Mori, M
中科院分区：
文献类型：
--
作者：
Mimori, K;Kataoka, A;Mori, M
比较正常细胞（N）、原发癌细胞（T）和转移癌细胞（M）之间的差异基因表达谱，可以确定与乳腺癌进展和转移直接相关的关键基因。采用激光显微切割技术（LMD）从6例T、N、M共20个切片中提取总RNA。经T7系统扩增后，用cDNA微阵列鉴定T、N和M之间的差异表达基因。此外，为了阐明基因表达改变的机制，我们通过亚硫酸氢盐处理后测序确定了有趣基因的甲基化状态。结果表明，与T相比，M中运动相关蛋白-1（MRP-1/CD 9）、外周髓鞘蛋白-22（PMP-22）和半胱天冬酶-3（CASP-3）的表达下调。对56例乳腺癌组织中MRP-1的表达进行了研究，发现MRP-1的表达与乳腺癌的分期呈负相关（P < 0.05），MRP-1阳性者5年无复发生存率明显高于阴性者（P < 0.05）。相反，与N相比，在T中观察到11倍的信号转导和翻译因子的过表达。在一例病例中仅在CASP-3中观察到癌症特异性甲基化。总之，本试验的建立使我们能够检测与肿瘤组织内每个细胞群直接相关的基因，并为我们提供了在临床乳腺癌病例中全面鉴定转移相关基因的线索。
Comparing differential gene expression profiles established by cDNA microarray between normal cells (N), primary carcinoma cells (T), and metastatic carcinoma cells (M) may determine those critical genes directly associated with progression and metastasis of breast cancer. Total RNA was extracted by laser microdissection (LMD) from 20 slices of T, N and M from 6 cases. After amplification by a T7-based system, differentially expressed genes between T, N and M were identified by cDNA microarray. In addition, to clarify the mechanism for altered gene expression, we determined the methylation status by sequencing after bisulfite treatment for intriguing genes. As a result, the expression of motility related protein-1 (MRP-1/CD9), peripheral myelin protein-22 (PMP-22), and caspase 3 (CASP-3) were down-regulated in M compared to T. We focused especially on MRP-1 and found that the expression status of MRP-1 was significantly inversely associated with stage of disease in 56 cases of breast cancer (P < 0.05), and the relapse free survival in 5 years was significantly higher in MRP-1 positive cases than those negative cases (P < 0.05). Conversely, overexpression, by 11-fold, of signal transduction and translation factors were observed in T compared to N. The cancer specific methylation was observed only in CASP-3 in a case. In conclusion, the establishment of the present assay allows us to detect genes directly associated with each cell population within tumor tissue and gives us clues to identify metastasis-related genes comprehensively in clinical breast cancer cases.