The State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610041, PR China.
Mol Cell Proteomics. 2011 Oct;10(10):M110.005397. doi: 10.1074/mcp.M110.005397. Epub 2011 May 27.
Colorectal cancer (CRC) is the third most commonly diagnosed cancer in males and second in females worldwide. Unfortunately 40-50% of patients already have metastatic disease at presentation when prognosis is poor with a 5-year survival of <10%. Reactive oxygen species (ROS) have been proposed to play a crucial role in tumor metastasis. We now show that higher levels of ROS accumulation are found in a colorectal cancer-derived metastatic cell line (SW620) compared with a cell line (SW480) derived from the primary lesion from the same patient. In addition, ROS accumulation can affect both the migratory and invasive capacity of SW480 and SW620 cells. To explore the molecular mechanism underlying ROS-induced migration and invasion in CRC, we have compared protein expression patterns between SW480 and SW620 cells using a two-dimensional electrophoresis-based proteomics strategy. A total of 63 altered proteins were identified from tandem MS analysis. Cluster analysis revealed dysregulated expression of multiple redox regulative or ROS responsive proteins, implicating their functional roles in colorectal cancer metastasis. Molecular and pathological validation demonstrated that altered expression of PGAM1, GRB2, DJ-1, ITGB3, SOD-1, and STMN1 was closely correlated with the metastatic potential of CRC. Functional studies showed that ROS markedly up-regulated expression of ITGB3, which in turn promoted an aggressive phenotype in SW480 cells, with concomitant up-regulated expression of STMN1. In contrast, knockdown of ITGB3 expression could mitigate the migratory and invasive potential of SW620 or H(2)O(2)-treated SW480 cells, accompanied by down-regulated expression of STMN1. The function of ITGB3 was dependent on the surface expression of integrin αvβ3 heterodimer. Furthermore, STMN1 expression and the PI3K-Akt-mTOR pathway were found to be involved in ROS-induced and ITGB3-mediated migration and invasion of colorectal cancer cells. Taken together, these studies suggest that ITGB3 plays an important role in ROS-induced migration and invasion in CRC.
结直肠癌（CRC）是全球男性中第三常见的癌症，女性中第二常见的癌症。不幸的是，40-50%的患者在就诊时已经患有转移性疾病，预后较差，5 年生存率<10%。活性氧（ROS）已被提出在肿瘤转移中起关键作用。我们现在表明，在源自同一患者原发性病变的细胞系（SW480）中发现的 ROS 积累水平低于源自转移性细胞系（SW620）的 ROS 积累水平。此外，ROS 积累可以影响 SW480 和 SW620 细胞的迁移和侵袭能力。为了探讨 CRC 中 ROS 诱导的迁移和侵袭的分子机制，我们使用基于二维电泳的蛋白质组学策略比较了 SW480 和 SW620 细胞之间的蛋白质表达模式。通过串联 MS 分析鉴定了 63 个改变的蛋白质。聚类分析显示，多个氧化还原调节或 ROS 反应蛋白的表达失调，暗示它们在结直肠癌转移中的功能作用。分子和病理学验证表明，PGAM1、GRB2、DJ-1、ITGB3、SOD-1 和 STMN1 的表达改变与 CRC 的转移潜能密切相关。功能研究表明，ROS 显著上调了 ITGB3 的表达，从而促进了 SW480 细胞的侵袭表型，同时上调了 STMN1 的表达。相反，下调 ITGB3 的表达可以减轻 SW620 或 H2O2 处理的 SW480 细胞的迁移和侵袭潜力，同时下调 STMN1 的表达。ITGB3 的功能依赖于整合素 αvβ3 异二聚体的表面表达。此外，发现 STMN1 表达和 PI3K-Akt-mTOR 途径参与了 ROS 诱导和 ITGB3 介导的结直肠癌细胞迁移和侵袭。总之，这些研究表明，ITGB3 在 CRC 中 ROS 诱导的迁移和侵袭中起重要作用。
