Section of Hematology/Oncology, Department of Medicine, Section of Urology, University of Chicago, Chicago, Illinois, USA.
Clin Cancer Res. 2011 May 15;17(10):3112-22. doi: 10.1158/1078-0432.CCR-10-1264. Epub 2011 Mar 1.
Fyn is a member of the Src family of kinases that we have previously shown to be overexpressed in prostate cancer. This study defines the biological impact of Fyn inhibition in cancer using a PC3 prostate cancer model.
Fyn expression was suppressed in PC3 cells using an shRNA against Fyn (PC3/FYN-). Knockdown cells were characterized using standard growth curves and time-lapse video microscopy of wound assays and Dunn Chamber assays. Tissue microarray analysis was used to verify the physiologic relevance of the HGF/MET axis in human samples. Flank injections of nude mice were performed to assess in vivo growth characteristics.
HGF was found to be sufficient to drive Fyn-mediated events. Compared to control transductants (PC3/Ctrl), PC3/FYN- showed a 21% decrease in growth at 4 days (P = 0.05). PC3/FYN- cells were 34% longer than control cells (P = 0.018) with 50% increase in overall surface area (P < 0.001). Furthermore, when placed in a gradient of HGF, PC3/FYN- cells showed impaired directed chemotaxis down an HGF gradient in comparison to PC3/Ctrl (P = 0.001) despite a 41% increase in cellular movement speed. In vivo studies showed 66% difference of PC3/FYN- cell growth at 8 weeks using bidimensional measurements (P = 0.002).
Fyn plays an important role in prostate cancer biology by facilitating cellular growth and by regulating directed chemotaxis-a key component of metastasis. This finding bears particular translational importance when studying the effect of Fyn inhibition in human subjects.
Fyn 是 Src 家族激酶的成员，我们之前已经证明其在前列腺癌中过度表达。本研究使用 PC3 前列腺癌细胞模型定义了 Fyn 抑制在癌症中的生物学影响。
使用针对 Fyn 的 shRNA（PC3/FYN-）抑制 PC3 细胞中的 Fyn 表达。使用标准生长曲线和划痕试验和 Dunn 室试验的延时视频显微镜对敲低细胞进行表征。组织微阵列分析用于验证 HGF/MET 轴在人类样本中的生理相关性。进行裸鼠的侧腹注射以评估体内生长特征。
发现 HGF 足以驱动 Fyn 介导的事件。与对照转染体（PC3/Ctrl）相比，PC3/FYN-在第 4 天的生长速度降低了 21%（P = 0.05）。PC3/FYN-细胞比对照细胞长 34%（P = 0.018），总表面积增加 50%（P < 0.001）。此外，当置于 HGF 梯度中时，与 PC3/Ctrl 相比，PC3/FYN-细胞的定向趋化性受损，沿 HGF 梯度向下迁移（P = 0.001），尽管细胞运动速度增加了 41%。体内研究表明，使用二维测量，8 周时 PC3/FYN-细胞生长差异为 66%（P = 0.002）。
Fyn 通过促进细胞生长和调节定向趋化性（转移的关键组成部分）在前列腺癌生物学中发挥重要作用。当研究 Fyn 抑制对人类受试者的影响时，这一发现具有特殊的转化意义。
