Barbolina Maria V, Adley Brian P, Kelly David L, Fought Angela J, Scholtens Denise M, Shea Lonnie D, Stack M Sharon
Department of Chemical and Biochemical Engineering, Northwestern University, Chicago, IL, USA.
Lab Invest. 2008 Jun;88(6):602-14. doi: 10.1038/labinvest.2008.25. Epub 2008 Mar 24.
Advanced and metastatic ovarian cancer is a leading cause of death from gynecologic malignancies. A more detailed understanding of the factors controlling invasion and metastasis may lead to novel anti-metastatic therapies. To model cellular interactions that occur during intraperitoneal metastasis, comparative cDNA microarray analysis and confirmatory real-time reverse transcription PCR (RT-PCR) were employed to uncover changes in gene expression that may occur in late stage ovarian cancer in response to microenvironmental cues, particularly native three-dimensional collagen I. Gene expression in human ovarian carcinoma tissues was evaluated on the RNA and protein level using real-time RT-PCR and immunohistochemistry. Cell invasion and migration were evaluated in a collagen invasion assay and a scratch wound assay. Three-dimensional collagen I culture led to differential expression of several genes. The role of actinin alpha-4 (ACTN4), a cytoskeleton-associated protein implicated in the regulation of cell motility, was examined in detail. ACTN4 RNA and protein expression were associated with advanced and metastatic human ovarian carcinoma. This report demonstrates that a cytoskeletal-associated protein ACTN4 is upregulated by three-dimensional collagen culture conditions, leading to increased invasion and motility of ovarian cancer cells. Expression of ACTN4 in human ovarian tumors was found to be associated with advanced-stage disease and peritoneal metastases.
晚期和转移性卵巢癌是妇科恶性肿瘤导致死亡的主要原因。对控制侵袭和转移的因素有更详细的了解可能会带来新的抗转移疗法。为了模拟腹膜内转移过程中发生的细胞相互作用，采用比较性cDNA微阵列分析和验证性实时逆转录PCR（RT-PCR）来揭示晚期卵巢癌可能因微环境线索，特别是天然三维I型胶原而发生的基因表达变化。使用实时RT-PCR和免疫组织化学在RNA和蛋白质水平评估人卵巢癌组织中的基因表达。在胶原侵袭试验和划痕伤口试验中评估细胞侵袭和迁移。三维I型胶原培养导致几个基因的差异表达。详细研究了肌动蛋白α-4（ACTN4）的作用，这是一种与细胞骨架相关的蛋白质，参与细胞运动的调节。ACTN4 RNA和蛋白质表达与晚期和转移性人卵巢癌相关。本报告表明，细胞骨架相关蛋白ACTN4在三维胶原培养条件下上调，导致卵巢癌细胞的侵袭和运动增加。发现ACTN4在人卵巢肿瘤中的表达与晚期疾病和腹膜转移相关。
