Boueroy Parichart, Saensa‑Ard Sunitta, Siripong Pongpun, Kanthawong Sakawrat, Hahnvajanawong Chariya
Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Center of Excellence for Innovation in Chemistry, Cholangiocarcinoma Research Institute, Cholangiocarcinoma Screening and Care Program (CASCAP), Faculty of Medicine, Khon Kaen University, Khon Kaen,Thailand. Email:
Asian Pac J Cancer Prev. 2018 Dec 25;19(12):3605-3613. doi: 10.31557/APJCP.2018.19.12.3605.
Cholangiocarcinoma is a malignant tumor with high metastatic and mortality rates. We investigated the effects of rhinacanthin-C on cell proliferation, migration, invasion and the expression of proteins regulating cancer cell invasion-regulated proteins in a cholangiocarcinoma (KKU-M156) cell line. Cytotoxicity of rhinacanthin-C was determined by the SRB assay. Using wound-migration, chamber-migration and chamber-invasion assays, we assessed the effects of rhinacanthin-C against KKU-M156 cells. The activities of matrix metalloproteinases 2 and 9 (MMP-2, MMP-9) and urokinase-type plasminogen activator (uPA) were determined using gelatinase and uPA zymography assays. The expression of invasion-regulated proteins was investigated using western-blot analysis. After treatment with rhinacanthin-C, KKU-M156 cells exhibited antiproliferative effects in a dose-dependent manner with greater efficacy than in Vero cells: IC50 values were 1.50 and 2.37 μM, respectively. Rhinacanthin-C significantly inhibited cell migration and invasion of KKU-M156 cells in a dose-dependent manner. Consistent with this observation, treatment with rhinacanthin-C was associated with a decrease in the expression levels of FAK, p-FAK, MMP-2, and a decrease in the levels of p38-, JNK1/2- and ERK1/2-MAPK pathways as well as inhibiting NF-κB/p65 expression and translocation of NF-κB/p65 to the nucleus. We have shown for the first time that the anti-metastatic effects of rhinacanthin-C on KKU-M156 cells are mediated via inhibition of the expression of invasion-regulated proteins. Rhinacanthin-C may deserve consideration as a potential agent for the treatment of cholangiocarcinoma.
胆管癌是一种具有高转移率和高死亡率的恶性肿瘤。我们研究了去甲氧基莪术二酮对胆管癌细胞系（KKU-M156）的细胞增殖、迁移、侵袭以及调控癌细胞侵袭相关蛋白表达的影响。采用SRB法测定去甲氧基莪术二酮的细胞毒性。通过伤口迁移试验、小室迁移试验和小室侵袭试验，评估去甲氧基莪术二酮对KKU-M156细胞的作用。使用明胶酶和尿激酶型纤溶酶原激活剂（uPA）酶谱分析法测定基质金属蛋白酶2和9（MMP-2、MMP-9）以及uPA的活性。采用蛋白质印迹分析研究侵袭相关蛋白的表达。用去甲氧基莪术二酮处理后，KKU-M156细胞呈现出剂量依赖性的抗增殖作用，且效果优于对非洲绿猴肾细胞（Vero细胞）：半数抑制浓度（IC50）值分别为1.50 μM和2.37 μM。去甲氧基莪术二酮以剂量依赖性方式显著抑制KKU-M156细胞的迁移和侵袭。与该观察结果一致，用去甲氧基莪术二酮处理会导致粘着斑激酶（FAK）、磷酸化FAK（p-FAK）、MMP-2的表达水平降低，同时p38丝裂原活化蛋白激酶（MAPK）、应激活化蛋白激酶1/2（JNK1/2）和细胞外信号调节激酶1/2（ERK1/2）-MAPK信号通路水平降低，以及抑制核因子κB（NF-κB）/p65的表达并阻止NF-κB/p65向细胞核的转位。我们首次表明，去甲氧基莪术二酮对KKU-M156细胞的抗转移作用是通过抑制侵袭相关蛋白的表达介导的。去甲氧基莪术二酮可能值得作为治疗胆管癌的潜在药物加以考虑。
