Division of Advanced Medical Science, Kobe University Graduate School of Science, Technology and Innovation, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, 650-0017, Japan.
Department of Public Health, Kobe University Graduate School of Health Sciences, 7-10-2 Tomogaoka, Suma-ku, Kobe, 654-0142, Japan.
J Cancer Res Clin Oncol. 2019 Sep;145(9):2261-2271. doi: 10.1007/s00432-019-02987-z. Epub 2019 Jul 31.
To investigate the role of sonic hedgehog (Shh) signaling and epithelial-mesenchymal transition (EMT) in bladder cancer progression and invasion.
We cultured three bladder cancer cell lines, muscle-invasive T24 and 5637, and non-muscle-invasive KK47, in the presence of a recombinant-Shh (r-Shh) protein or cyclopamine, a Shh signaling inhibitor, to investigate proliferation and expression of EMT markers. Wound-healing assays and transwell assay were performed to evaluate cell invasion and migration. Mice were then inoculated with bladder cancer cells and treated with cyclopamine. Mouse tumor samples were stained for Shh signaling and EMT markers.
R-Shh protein enhanced cell proliferation, whereas cyclopamine significantly suppressed cell proliferation, especially in invasive cancer (5637 and T24) (p < 0.05). R-Shh protein promoted EMT, suppressed E-cadherin and enhanced N-cadherin and vimentin and Gli1, an Shh downstream molecule, while cyclopamine blocked EMT, especially in 5637 and T24. Cyclopamine also inhibited cell invasion and migration in vitro. In the animal study, intraperitoneal injection of cyclopamine significantly suppressed tumor growth in 5637 and T24 in mice (p = 0.01 and p = 0.004, respectively) and slightly suppressing KK47 tumor growth (p = 0.298). Significant cyclopamine-induced suppression of Gli1 in 5637 and T24 mouse tumors (both p = 0.03) was seen, suggesting that muscle-invasive bladder cancer may be more dependent on Shh signaling than non-muscle-invasive bladder cancer.
Shh signaling and EMT were especially enhanced in muscle-invasive bladder cancer progression and invasion, and suppressed by the inhibition of Shh signaling.
研究 sonic hedgehog（Shh）信号通路和上皮间质转化（EMT）在膀胱癌进展和侵袭中的作用。
我们培养了三种膀胱癌细胞系，肌肉浸润性 T24 和 5637 以及非肌肉浸润性 KK47，在重组-Shh（r-Shh）蛋白或 Shh 信号通路抑制剂 cyclopamine 的存在下，研究 EMT 标志物的增殖和表达。进行划痕愈合试验和 Transwell 试验评估细胞侵袭和迁移。然后用膀胱癌细胞接种小鼠并给予 cyclopamine 治疗。对小鼠肿瘤样本进行 Shh 信号通路和 EMT 标志物染色。
r-Shh 蛋白增强细胞增殖，而 cyclopamine 则显著抑制细胞增殖，尤其是在侵袭性癌症（5637 和 T24）中（p<0.05）。r-Shh 蛋白促进 EMT，抑制 E-钙黏蛋白，增强 N-钙黏蛋白和波形蛋白，以及 Shh 下游分子 Gli1，而 cyclopamine 阻断 EMT，尤其是在 5637 和 T24 中。cyclopamine 还抑制体外细胞侵袭和迁移。在动物研究中，腹腔注射 cyclopamine 显著抑制了 5637 和 T24 小鼠肿瘤的生长（p=0.01 和 p=0.004，分别），并轻微抑制了 KK47 肿瘤的生长（p=0.298）。在 5637 和 T24 小鼠肿瘤中观察到 cyclopamine 诱导的 Gli1 显著抑制（均 p=0.03），这表明肌肉浸润性膀胱癌可能比非肌肉浸润性膀胱癌更依赖 Shh 信号通路。
Shh 信号通路和 EMT 在肌肉浸润性膀胱癌的进展和侵袭中明显增强，而 Shh 信号通路的抑制则会受到抑制。
